Inhibition of bacterial biofilms with imidazole-phenyl derivatives

ABSTRACT

Disclosure is provided for imidazole-phenyl derivative compounds that prevent, remove and/or inhibit the formation of biofilms, compositions comprising these compounds, devices comprising these compounds, and methods of using the same.

RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.12/417,981, filed Apr. 3, 2009, now allowed, and claims the benefitunder 35 U.S.C. § 119(e) of U.S. Application No. 61/042,473, filed Apr.4, 2008, the disclosure of which is incorporated herein by reference inits entirety. This application is related to U.S. application Ser. No.12/020,112, filed Jan. 25, 2008, and published Jul. 31, 2008, aspublication no. 2008/0181923, the disclosure of which is incorporatedherein by reference in its entirety.

FIELD OF THE INVENTION

This invention relates to compounds, compositions and methods useful forcontrolling biofilms and microorganisms.

BACKGROUND OF THE INVENTION

Biofilms are complex communities of microorganisms that are commonlyfound on a variety of substrates or surfaces that are moist or submerged(Musk et al., Curr. Med. Chem., 2006, 13, 2163; Donlan et al., Clin.Microbiol. Rev., 2002, 15, 167). Though primarily populated by bacteria,biofilms can also contain many different individual types ofmicroorganisms, e.g., bacteria, archaea, protozoa and algae. Theformation of biofilms can be thought of as a developmental process inwhich a few free-swimming (planktonic) bacteria adhere to a solidsurface and, in response to appropriate signals, initiate the formationof a complex sessile microcolony existing as a community of bacteria andother organisms. Bacteria within biofilms are usually embedded within amatrix, which can consist of protein, polysaccharide, nucleic acids, orcombinations of these macromolecules. The matrix is a critical featureof the biofilm that protects the inhabiting organisms from antiseptics,microbicides, and host cells. It has been estimated that bacteria withinbiofilms are upwards of 1,000-fold more resistant to conventionalantibiotics (Rasmussen et al., Int. J. Med. Microbiol., 2006, 296, 149).

Biofilms play a significant role in infectious disease. It is estimatedthat biofilms account for between 50-80% of microbial infections in thebody, and that the cost of these infections exceeds $1 billion annually.For example, persistent infections of indwelling medical devices remaina serious problem for patients, because eradication of these infectionsis virtually impossible. A few diseases in which biofilms have beenimplicated include endocarditis, otitis media, chronic prostatitis,periodontal disease, chronic urinary tract infections, and cysticfibrosis. The persistence of biofilm populations is linked to theirinherent insensitivity to antiseptics, antibiotics, and otherantimicrobial compounds or host cells.

Deleterious effects of biofilms are also found in non-medical settings.For example, biofilms are a major problem in the shipping industry.Biofilms form on and promote the corrosion of ship hulls and alsoincrease the roughness of the hulls, increasing the drag on the shipsand thereby increasing fuel costs. The biofilms can also promote theattachment of larger living structures, such as barnacles, to the hull.Fuel can account for half of the cost of marine shipping, and the lossin fuel efficiency due to biofilm formation is substantial. One methodof controlling biofilms is to simply scrape the films off of the hulls.However, this method is costly and time-consuming, and can promote thespread of troublesome non-native species in shipping waters. Anothermethod involves the use of antifouling coatings containing tin. However,tin-based coatings are now disfavored due to toxicity concerns.

Given the breadth of detrimental effects caused by bacterial biofilms,there has been an effort to develop small molecules that will inhibittheir formation (Musk et al., Curr. Med Chem., 2006, 13, 2163). Theunderlying principle is that if bacteria can be maintained in theplanktonic state, they will either not attach to a target surface and/orthey can be killed by a lower dose of microbicide.

Despite the extent of biofilm driven problems, examples of structuralscaffolds that inhibit biofilm formation are rare (Musk et al., Curr.Med. Chem., 2006, 13, 2163). The few known examples include thehomoserine lactones (Geske et al., J. Am. Chem. Soc., 2005, 127, 12762),which are naturally-occurring bacterial signaling molecules thatbacteria use in quorum sensing (Dong et al., J. Microbiol., 2005, 43,101; Nealson et al., J. Bacteria, 1970, 104, 313), brominated furanonesisolated from the macroalga Delisea pulchra (Hentzer et al.,Microbiology-Sgm, 2002, 148, 87), and ursene triterpenes from the plantDiospyros dendo (Hu et al., J. Nat. Prod., 2006, 69, 118).

Bacteria have an unparalleled ability to overcome foreign chemicalinsult. For example, resistance to vancomycin, “the antibiotic of lastresort,” has become more prevalent, and strains of vancomycin-resistantStaphylococcus aureus have become a serious health risk. It has beenpredicted that it is simply a matter of time before different bacterialstrains develop vancomycin resistance, and the safety net thatvancomycin has provided for decades in antibiotic therapy will no longerbe available.

Because of their natural resistance to antibiotics, phagocytic cells,and other biocides, biofilms are difficult, if not impossible, toeradicate. Therefore, the identification of compounds that controlbiofilm formation is of critical need.

SUMMARY OF THE INVENTION

Provided herein are compounds of Formula (I):

wherein:

R¹, R^(1′), R², R^(2′) and R³ are each independently selected from thegroup consisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide;

A and B are each independently selected from N, S and O; and

n=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (I), R¹ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (I), R¹ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (I), R³ is an amino and A and B are eachN, depicted as Formula (I)(a):

wherein:

R¹, R^(1′), R² and R^(2′) are each independently selected from the groupconsisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, sulfone, sulfoxide,oxo, oxy, nitro, carbonyl, carboxy, amino acid sidechain, amino acid andpeptide; and

n=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (I)(a), R¹ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (I)(a), R¹ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

Also provided are compounds of Formula (II):

wherein:

R⁴, R^(4′), R⁵, R^(5′) and R⁶ are each independently selected from thegroup consisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide;

A and B are each independently selected from N; S and O; and

n=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (II), R⁴ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (II), R⁴ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (II), R⁶ is an amino and A and B are eachN, depicted as Formula (II)(a):

wherein:

R⁴, R^(4′), R⁵ and R^(5′) are each independently selected from the groupconsisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, sulfone, sulfoxide,oxo, oxy, nitro, carbonyl, carboxy, amino acid sidechain, amino acid andpeptide;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (II)(a), R⁴ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (II)(a), R⁴ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

Further provided are compounds of Formula (III):

wherein:

R⁹, R^(9′), R¹⁰, R^(10′) and R¹¹ are each independently selected fromthe group consisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide;

A and B are each independently selected from N, S and O;

D, E, F, G, H and J are each independently selected from C, N, S and O,wherein at least one of D, E, F, G, H and J is C; and

n=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments of Formula (III), R⁹ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (III), R⁹ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

Also provided are compounds of Formula (IV):

wherein:

R¹², R^(12′), R¹³, R¹⁴ and R^(14′) are each independently selected fromthe group consisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide;

A and B are each independently selected from N, S and O; and

q=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (IV), R¹³ is an amino and A and B areeach N, depicted as Formula (IV)(a):

wherein:

R¹², R^(12′), R¹⁴ and R^(14′) are each independently selected from thegroup consisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide; and

q=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

Embodiments of Formula (IV)(a) include:

wherein:

R¹⁴ is selected from the group consisting of H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

q=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (IV)(a), R¹⁴ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (IV)(a), R¹⁴ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

Further provided are compounds of Formula (V):

wherein:

R¹⁵, R^(15′), R¹⁶, R^(16′), R¹⁷ and R¹⁸ are each independently selectedfrom the group consisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide;

A and B are each independently selected from N, S and O;

n=0 to 20, saturated or unsaturated; and

p=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (V), R¹⁸ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (V), R¹⁸ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (V), R¹⁷ is an amino and A and B are eachN, depicted as Formula (V)(a):

wherein:

R¹⁵, R^(15′), R¹⁶, R^(16′) and R¹⁸ are each independently selected fromthe group consisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide;

n=0 to 20, saturated or unsaturated; and

p=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (V)(a), R¹⁸ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (V)(a), R¹⁸ is a group:

wherein:

R⁸ is selected from the group consisting of H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (V)(a), n=0 and R¹⁵ and R^(15′) are eachH, depicted as Formula (V)(a)(1):

wherein:

R¹⁶, R^(16′) and R¹⁸ are each independently selected from the groupconsisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol, sulfone,sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acid sidechain,amino acid and peptide; and

p=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (V)(a)(1), R¹⁸ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (V)(a)(1), R¹⁸ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

Biofilm preventing, removing or inhibiting compositions are provided,which include a carrier and an effective amount of a compound disclosedherein. In some embodiments, the composition is a dentifrice compositionthat promotes dental hygiene by preventing, reducing, inhibiting orremoving a biofilm. In some embodiments, the dentifrice compositioncomprises a toothpaste, mouthwash, chewing gum, dental floss, or dentalcream.

Compositions are also provided that include a compound disclosed hereinin a pharmaceutically acceptable carrier.

Compositions are further provided that include a compound disclosedherein covalently coupled to a substrate. In some embodiments, thesubstrate includes a polymeric material. In some embodiments, thesubstrate includes a solid support. In some embodiments, the substrateincludes a drainpipe, glaze ceramic, porcelain, glass, metal, wood,chrome, plastic, vinyl, and Formica® brand laminate (The DillerCorporation, Cincinnati, Ohio). In some embodiments, the substrateincludes shower curtains or liners, upholstery, laundry, and carpeting.In some embodiments, the substrate includes a ship hull or a portionthereof. In some embodiments, the substrate includes a food contactsurface.

Biofilm preventing, removing or inhibiting coating compositions areprovided, including: (a) a film-forming resin; (b) a solvent thatdisperses said resin; (c) an effective amount of the compounds orcompositions disclosed herein, wherein said effective amount prevents orinhibits the growth of a biofilm thereon; and (d) optionally, at leastone pigment. In some embodiments, the compound is covalently coupled tothe resin. In some embodiments, the resin includes a polymeric material.

Substrates coated with coating composition disclosed herein are alsoprovided. In some embodiments, the substrate includes a polymericmaterial. In some embodiments, the substrate includes a solid support.In some embodiments, the substrate includes a drainpipe, glaze ceramic,porcelain, glass, metal, wood, chrome, plastic, vinyl, and Formica®brand laminate. In some embodiments, the substrate includes showercurtains or liners, upholstery, laundry, and carpeting. In someembodiments, the substrate includes a ship hull or a portion thereof. Insome embodiments, the substrate includes a food contact surface.

Methods of controlling biofilm formation on a substrate are provided,including the step of contacting the substrate with a compound and/orcomposition disclosed herein in an amount effective to inhibit biofilmformation. In some embodiments, controlling biofilm formation includesclearing a preformed biofilm from said substrate by administering aneffective amount of the compound and/or composition disclosed herein tosaid substrate, wherein said effective amount will reduce the amount ofsaid biofilm on said substrate. In some embodiments, the substrate mayinclude a drainpipe, glaze ceramic, porcelain, glass, metal, wood,chrome, plastic, vinyl, and Formica® brand laminate. In someembodiments, the substrate may include a food product (e.g., seafood).In some embodiments, the biofilm includes Gram-negative bacteria.

Methods for treating and/or preventing a bacterial infection (e.g.,chronic bacterial infection) in a subject in need thereof are provided,including administering to said subject a compound and/or compositiondisclosed herein in an amount effective to inhibit, reduce, or remove abiofilm component of said bacterial infection. The bacterial infectionmay include urinary tract infection, gastritis, respiratory infection,cystitis, pyelonephritis, osteomyelitis, bacteremia, skin infection,rosacea, acne, chronic wound infection, infectious kidney stones,bacterial endocarditis, and sinus infection.

Also provided are medical devices, including (a) a medical devicesubstrate; and (b) an effective amount of a compound disclosed herein,either coating the substrate, or incorporated into the substrate,wherein said effective amount prevents or inhibits the growth of abiofilm thereon. In some embodiments, the medical device substrate mayinclude stents, fasteners, ports, catheters, scaffolds and grafts. Insome embodiments, the compound is covalently coupled to said substrate.

Compounds and/or compositions for use in a method to control a biofilmare further provided. Also provided is the use of compounds and/orcompositions disclosed herein for the preparation of a medicament forthe treatment and/or prevention of a bacterial infection (e.g., chronicbacterial infection).

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The present invention is further described below. All U.S. patentreferences referred to in this patent application are herebyincorporated by reference in their entirety as if set forth fullyherein.

A. Definitions

“Imidazole” refers to the commonly known structure:

“H” refers to a hydrogen atom. “C” refers to a carbon atom. “N” refersto a nitrogen atom. “O” refers to an oxygen atom. “Halo” refers to F,Cl, Br or I. The term “hydroxy,” as used herein, refers to an —OHmoiety. “Br” refers to a bromine atom. “Cl” refers to a chlorine atom.“I” refers to an iodine atom. “F” refers to a fluorine atom.

An “acyl group” is intended to mean a —C(O)—R radical, where R is asuitable substituent (for example, an acetyl group, a propionyl group, abutyroyl group, a benzoyl group, or an alkylbenzoyl group).

“Alkyl,” as used herein, refers to a straight or branched chainhydrocarbon containing from 1 or 2 to 10 or 20 or more carbon atoms(e.g., C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15,etc.). Representative examples of alkyl include, but are not limited to,methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl,tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl,2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl,n-decyl, and the like. In some embodiments, alkyl groups as describedherein are optionally substituted (e.g., from 1 to 3 or 4 times) withindependently selected H, halo, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide.

The term “optionally substituted” indicates that the specified group iseither unsubstituted, or substituted by one or more suitablesubstituents. A “substituent” is an atom or atoms substituted in placeof a hydrogen atom on the parent chain or cycle of an organic molecule,for example, H, hydroxy, acyl, alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol, sulfone,sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acid sidechain,amino acid and peptide.

“Alkenyl,” as used herein, refers to a straight or branched chainhydrocarbon containing from 1 or 2 to 10 or 20 or more carbons, andcontaining at least one carbon-carbon double bond, formed structurally,for example, by the replacement of two hydrogens. Representativeexamples of “alkenyl” include, but are not limited to, ethenyl,2-propenyl, 2-methyl-2-propenyl, 3-butenyl, 4-pentenyl, 5-hexenyl,2-heptenyl, 2-methyl-1-heptenyl, 3-decenyl and the like. In someembodiments, alkenyl groups as described herein are optionallysubstituted (e.g., from 1 to 3 or 4 times) with independently selectedH, halo, hydroxy, acyl, alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol, sulfone,sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acid sidechain,amino acid and peptide.

“Alkynyl,” as used herein, refers to a straight or branched chainhydrocarbon group containing from 1 or 2 to 10 or 20 or more carbonatoms, and containing at least one carbon-carbon triple bond.Representative examples of alkynyl include, but are not limited, toacetylenyl, 1-propynyl, 2-propynyl, 3-butynyl, 2-pentynyl, 1-butynyl andthe like. In some embodiments, alkynyl groups as described herein areoptionally substituted (e.g., from 1 to 3 or 4 times) with independentlyselected H, halo, hydroxy, acyl, alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol, sulfone,sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acid sidechain,amino acid and peptide.

The term “cycloalkyl,” as used herein, refers to a saturated cyclichydrocarbon group containing from 3 to 8 carbons or more. Representativeexamples of cycloalkyl include, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, and cyclooctyl. In some embodiments, cycloalkylgroups as described herein are optionally substituted (e.g., from 1 to 3or 4 times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

“Heterocyclo,” as used herein, refers to a monocyclic or a bicyclic ringsystem. Monocyclic heterocycle ring systems are exemplified by any 5 or6 member ring containing 1, 2, 3, or 4 heteroatoms independentlyselected from the group consisting of: O, N, and S. The 5 member ringhas from 0 to 2 double bonds, and the 6 member ring has from 0-3 doublebonds. Representative examples of monocyclic ring systems include, butare not limited to, azetidine, azepine, aziridine, diazepine,1,3-dioxolane, dioxane, dithiane, furan, imidazole, imidazoline,imidazolidine, isothiazole, isothiazoline, isothiazolidine, isoxazole,isoxazoline, isoxazolidine, morpholine, oxadiazole, oxadiazoline,oxadiazolidine, oxazole, oxazoline, oxazolidine, piperazine, piperidine,pyran, pyrazine, pyrazole, pyrazoline, pyrazolidine, pyridine,pyrimidine, pyridazine, pyrrole, pyrroline, pyrrolidine,tetrahydrofuran, tetrahydrothiophene, tetrazine, tetrazole, thiadiazole,thiadiazoline, thiadiazolidine, thiazole, thiazoline, thiazolidine,thiophene, thiomorpholine, thiomorpholine sulfone, sulfoxide, thiopyran,triazine, triazole, trithiane, and the like. Bicyclic ring systems areexemplified by any of the above monocyclic ring systems fused to an arylgroup as defined herein, a cycloalkyl group as defined herein, oranother monocyclic ring system as defined herein. Representativeexamples of bicyclic ring systems include but are not limited to, forexample, benzimidazole, benzothiazole, benzothiadiazole, benzothiophene,benzoxadiazole, benzoxazole, benzofuran, benzopyran, benzothiopyran,benzodioxine, 1,3-benzodioxole, cinnoline, indazole, indole, indoline,indolizine, naphthyridine, isobenzofuran, isobenzothiophene, isoindole,isoindoline, isoquinoline, phthalazine, pyranopyridine, quinoline,quinolizine, quinoxaline, quinazoline, tetrahydroisoquinoline,tetrahydroquinoline, thiopyranopyridine, and the like.

“Aryl” as used herein refers to a fused ring system having one or morearomatic rings. Representative examples of aryl include, azulenyl,indanyl, indenyl, naphthyl, phenyl, tetrahydronaphthyl, and the like.The aryl groups of this invention can be substituted with 1, 2, 3, 4, or5 substituents independently selected from alkenyl, alkenyloxy, alkoxy,alkoxyalkoxy, alkoxycarbonyl, alkyl, alkylcarbonyl, alkylcarbonyloxy,alkylsulfinyl, alkylsulfonyl, alkylthio, alkynyl, aryl, aryloxy, azido,arylalkoxy, arylalkyl, aryloxy, carboxy, cyano, formyl, halogen,haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, mercapto, nitro, sulfamyl,sulfo, sulfonate, —NR′R″ (wherein, R′ and R″ are independently selectedfrom hydrogen, alkyl, alkylcarbonyl, aryl, arylalkyl and formyl), and—C(O)NR′R″ (wherein R′ and R″ are independently selected from hydrogen,alkyl, alkylcarbonyl, aryl, arylalkyl, and formyl).

“Heteroaryl” means a cyclic, aromatic hydrocarbon in which one or morecarbon atoms have been replaced with heteroatoms. If the heteroarylgroup contains more than one heteroatom, the heteroatoms may be the sameor different. Examples of heteroaryl groups include pyridyl,pyrimidinyl, imidazolyl, thienyl, furyl, pyrazinyl, pyrrolyl, pyranyl,isobenzofuranyl, chromenyl, xanthenyl, indolyl, isoindolyl, indolizinyl,triazolyl, pyridazinyl, indazolyl, purinyl, quinolizinyl, isoquinolyl,quinolyl, phthalazinyl, naphthyridinyl, quinoxalinyl, isothiazolyl, andbenzo[b]thienyl. Preferred heteroaryl groups are five and six memberedrings and contain from one to three heteroatoms independently selectedfrom the group consisting of: O, N, and S. The heteroaryl group,including each heteroatom, can be unsubstituted or substituted with from1 to 4 suitable substituents, as chemically feasible. For example, theheteroatom S may be substituted with one or two oxo groups, which may beshown as ═O.

“Alkoxy,” as used herein, refers to an alkyl group, as defined herein,appended to the parent molecular moiety through an oxy group, as definedherein. Representative examples of alkoxy include, but are not limitedto, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, tert-butoxy, pentyloxy,hexyloxy and the like. In some embodiments, alkoxy groups as describedherein are optionally substituted (e.g., from 1 to 3 or 4 times) withindependently selected H, halo, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide.

An “amine” or “amino” group is intended to mean the group —NH₂.“Optionally substituted” amines refers to —NH₂ groups wherein none, oneor two of the hydrogens is replaced by a suitable substituent asdescribed herein, such as alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclo, aryl, heteroaryl, alkoxy, carbonyl, carboxy, etc. In someembodiments, one or two of the hydrogens are optionally substituted withindependently selected, halo, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide. Disubstituted amines may havesubstituents that are bridging, i.e., form a heterocyclic ring structurethat includes the amine nitrogen.

An “amide” as used herein refers to an organic functional group having acarbonyl group (C═O) linked to a nitrogen atom (N), or a compound thatcontains this group, generally depicted as:

wherein, R and R′ can independently be any covalently-linked atom oratoms, for example, H, halo, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide.

A “thiol” or “mercapto” refers to an —SH group or to its tautomer ═S.

A “sulfone” as used herein refers to a sulfonyl functional group,generally depicted as:

wherein, R can be any covalently-linked atom or atoms, for example, H,halo, hydroxy, acyl, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclo,aryl, heteroaryl, alkoxy, amino, amide, thiol, sulfone, sulfoxide, oxo,oxy, nitro, carbonyl, carboxy, amino acid sidechain, amino acid andpeptide.

A “sulfoxide” as used herein refers to a sulfinyl functional group,generally depicted as:

wherein, R can be any covalently-linked atom or atoms, for example, H,halohydroxy, acyl, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclo,aryl, heteroaryl, alkoxy, amino, amide, thiol, sulfone, sulfoxide, oxo,oxy, nitro, carbonyl, carboxy, amino acid sidechain, amino acid andpeptide.

“Triazole” refers to the commonly known structures:

The term “oxo,” as used herein, refers to a ═O moiety. The term “oxy,”as used herein, refers to a —O— moiety.

“Nitro” refers to the organic compound functional group —NO₂.

“Carbonyl” is a functional group having a carbon atom double-bonded toan oxygen atom (—C═O). “Carboxy” as used herein refers to a —COOHfunctional group, also written as —(C═O)—OH.

“Amino acid sidechain” as used herein refers to any of the 20 commonlyknown groups associated with naturally-occurring amino acids, or anynatural or synthetic homologue thereof. An “amino acid” includes thesidechain group and the amino group, alpha-carbon atom, and carboxygroups, as commonly described in the art. Examples of amino acidsinclude glycine, and glycine that is substituted with a suitablesubstituent as described herein, such as alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, carbonyl, carboxy,etc., or a pharmaceutically acceptable salt or prodrug thereof. Forexample, “Histidine” is one of the 20 most commonly known amino acidsfound naturally in proteins. It contains an imidazole side chainsubstituent. Other examples of naturally-occurring amino acids includelysine, arginine, aspartic acid, glutamic acid, asparagine, glutamine,serine, threonine, tyrosine, alanine, valine, leucine, isoleucine,phenylalanine, methionine, cryptophan, and cysteine. Also included inthe definitions of “amino acid sidechain” and “amino acid” is proline,which is commonly included in the definition of an amino acid, but istechnically an imino acid. As used in this application, both thenaturally-occurring L-, and the non-natural D-amino acid enantiomers areincluded. A “peptide” is a linear chain of amino acids covalently linkedtogether, typically through an amide linkage, and contains from 1 or 2to 10 or 20 or more amino acids, and is also optionally substitutedand/or branched.

“Boc” or “BOC” is t-butoxycarbonyl, a commonly-known amino protectinggroup.

A “pharmaceutically acceptable salt” is intended to mean a salt thatretains the biological effectiveness of the free acids and bases of aspecified compound and that is not biologically or otherwiseundesirable. Examples of pharmaceutically acceptable salts includesulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates,monohydrogenphosphates, dihydrogenphosphates, metaphosphates,pyrophosphates, chlorides, bromides, iodides, acetates, propionates,decanoates, caprylates, acrylates, formates, isobutyrates, caproates,heptanoates, propiolates, oxalates, malonates, succinates, suberates,sebacates, fumarates, maleates, butyne-1,4-dioates, hexyne-1,6-dioates,benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates,hydroxybenzoates, methoxybenzoates, phthalates, sulfonates,xylenesulfonates, phenylacetates, phenylpropionates, phenylbutyrates,citrates, lactates, γ-hydroxybutyrates, glycollates, tartrates,methane-sulfonates, propanesulfonates, naphthalene-1-sulfonates,naphthalene-2-sulfonates, and mandelates.

A “prodrug” is intended to mean a compound that is converted underphysiological conditions or by solvolysis or metabolically to aspecified compound that is pharmaceutically active. A thoroughdiscussion is provided in T. Higuchi and V. Stella, Prodrugs as Noveldelivery Systems, Vol. 14 of the A.C.S. Symposium Series and in EdwardB. Roche, ed., Bioreversible Carriers in Drug Design, AmericanPharmaceutical Association and Pergamon Press, 1987, both of which areincorporated by reference herein in their entirety.

B. Active Compounds

Active compounds are provided herein that are useful to prevent, removeand/or inhibit the formation of biofilms. In some embodiments, activecompounds are derivatives of imidazole. Active compounds as describedherein can be prepared as detailed below or in accordance with knownprocedures or variations thereof that will be apparent to those skilledin the art.

As will be appreciated by those of skill in the art, the activecompounds of the various formulas disclosed herein may contain chiralcenters, e.g. asymmetric carbon atoms. Thus, the present invention isconcerned with the synthesis of both: (i) racemic mixtures of the activecompounds, and (ii) enantiomeric forms of the active compounds. Theresolution of racemates into enantiomeric forms can be done inaccordance with known procedures in the art. For example, the racematemay be converted with an optically active reagent into a diastereomericpair, and the diastereomeric pair subsequently separated into theenantiomeric forms.

Geometric isomers of double bonds and the like may also be present inthe compounds disclosed herein, and all such stable isomers are includedwithin the present invention unless otherwise specified. Also includedin active compounds of the invention are tautomers (e.g., tautomers ofimidazole) and rotamers.

Active compounds invention include compounds of Formula (I):

wherein:

R¹, R^(1′), R², R^(2′) and R³ are each independently selected from thegroup consisting of H, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide;

A and B are each independently selected from N, S and O; and

n=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (I), R¹ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (I), R¹ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (I), R³ is an amino and A and B are eachN, depicted as Formula (I)(a):

wherein:

R¹, R^(1′), R² and R^(2′) are each independently selected from the groupconsisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, sulfone, sulfoxide,oxo, oxy, nitro, carbonyl, carboxy, amino acid sidechain, amino acid andpeptide; and

n=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (I)(a), R¹ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (I)(a), R¹ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

Active compounds include compounds of Formula (II):

wherein:

R⁴, R^(4′), R⁵, R^(5′) and R⁶ are each independently selected from thegroup consisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide;

A and B are each independently selected from N, S and O; and

n=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (II), R⁴ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (II), R⁴ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (II), R⁶ is an amino and A and B are eachN, depicted as Formula (II)(a):

wherein:

R⁴, R^(4′), R⁵ and R^(5′) are each independently selected from the groupconsisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, sulfone, sulfoxide,oxo, oxy, nitro, carbonyl, carboxy, amino acid sidechain, amino acid andpeptide;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (II)(a), R⁴ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (II)(a), R⁴ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

Active compounds also include compounds of Formula (III):

wherein:

R⁹, R^(9′), R¹⁰, R^(10′) and R¹¹ are each independently selected fromthe group consisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide;

A and B are each independently selected from N, S and O;

D, E, F, G, H and J are each independently selected from C, N, S and O,wherein at least one of D, E, F, G, H and J is C; and

n=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments of Formula (III), R⁹ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (III), R⁹ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

Active compounds include those of Formula (IV):

wherein:

R¹², R^(12′), R¹³, R¹⁴ and R^(14′) are each independently selected fromthe group consisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide;

A and B are each independently selected from N, S and O; and

q=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (IV), R¹³ is an amino and A and B areeach N, depicted as Formula (IV)(a):

wherein:

R¹², R^(12′), R¹⁴ and R^(14′) are each independently selected from thegroup consisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide; and

q=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

Embodiments of Formula (IV)(a) include:

wherein:

R¹⁴ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

q=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (IV)(a), R¹⁴ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (IV)(a), R¹⁴ is a group:

wherein:

R⁸ is selected from the group consisting of H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

Active compounds invention include compounds of Formula (V):

wherein:

R¹⁵, R^(15′), R¹⁶, R^(16′), R¹⁷ and R¹⁸ are each independently selectedfrom the group consisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide;

A and B are each independently selected from N, S and O;

n=0 to 20, saturated or unsaturated; and p=0 to 20, saturated orunsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (V), R¹⁸ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (V), R¹⁸ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (V), R¹⁷ is an amino and A and B are eachN, depicted as Formula (V)(a):

wherein:

R¹⁵, R^(15′), R¹⁶, R^(16′) and R¹⁸ are each independently selected fromthe group consisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol,sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acidsidechain, amino acid and peptide;

n=0 to 20, saturated or unsaturated; and

p=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (V)(a), R¹⁸ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (V)(a), R¹⁸ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (V)(a), n=0 and R¹⁵ and R^(15′) are eachH, depicted as Formula (V)(a)(1):

wherein:

R¹⁶, R^(16′) and R¹⁸ are each independently selected from the groupconsisting of: H, hydroxy, acyl, alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclo, aryl, heteroaryl, alkoxy, amino, amide, thiol, sulfone,sulfoxide, oxo, oxy, nitro, carbonyl, carboxy, amino acid sidechain,amino acid and peptide; and

p=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

This formula may be optionally substituted (e.g., from 1 to 3 or 4times) with independently selected H, halo, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, thiol, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl,carboxy, amino acid sidechain, amino acid and peptide.

In some embodiments, one, two or three of the carbon atoms in the phenylmoiety is replaced by a suitable heteroatom, e.g., with a nitrogen(e.g., pyridine, pyrimidine, pyrazine, etc., heterocycles), oxygen,sulfur, etc.

In some embodiments of Formula (V)(a)(1), R¹⁸ is a group:

wherein:

R⁷ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

In some embodiments of Formula (V)(a)(1), R¹⁸ is a group:

wherein:

R⁸ is selected from the group consisting of: H, hydroxy, acyl, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl, alkoxy,amino, amide, sulfone, sulfoxide, oxo, oxy, nitro, carbonyl, carboxy,amino acid sidechain, amino acid and peptide; and

m=0 to 20, saturated or unsaturated;

or a pharmaceutically acceptable salt or prodrug thereof.

C. Compositions

In some embodiments, biofilm preventing, removing or inhibitingcompositions are provided, comprising a carrier and an effective amountof active compound. “Biofilm” or “biofilms” refer to communities ofmicroorganisms that are attached to a substrate. The microorganismsoften excrete a protective and adhesive matrix of polymeric compounds.They often have structural heterogeneity, genetic diversity, and complexcommunity interactions. “Biofilm preventing”, “biofilm removing”,“biofilm inhibiting”, “biofilm reducing”, “biofilm resistant”, “biofilmcontrolling” or “antifouling” refer to prevention of biofilm formation,inhibition of the establishment or growth of a biofilm, or decrease inthe amount of organisms that attach and/or grow upon a substrate, up toand including the complete removal of the biofilm. As used herein, a“substrate” can include any living or nonliving structure. For example,biofilms often grow on synthetic materials submerged in an aqueoussolution or exposed to humid air, but they also can form as floatingmats on a liquid surface, in which case the microorganisms are adheringto each other or to the adhesive matrix characteristic of a biofilm. An“effective amount” of a biofilm preventing, removing or inhibitingcomposition is that amount which is necessary to carry out thecomposition's function of preventing, removing or inhibiting a biofilm.

In some embodiments, the carrier is a pharmaceutically acceptablecarrier. A “pharmaceutically acceptable carrier” as used herein refersto a carrier that, when combined with an active compound of the presentinvention, facilitates the application or administration of that activecompound for its intended purpose to prevent or inhibit biofilmformation, or remove an existing biofilm. The active compounds may beformulated for administration in a pharmaceutically acceptable carrierin accordance with known techniques. See, e.g., Remington, The ScienceAnd Practice of Pharmacy (9^(th) Ed. 1995). The pharmaceuticallyacceptable carrier must, of course, also be acceptable in the sense ofbeing compatible with any other ingredients in the composition. Thecarrier may be a solid or a liquid, or both, and is preferablyformulated with the compound as a unit-dose composition, for example, atablet, which may contain from 0.01 or 0.5% to 95% or 99% by weight ofthe active compound. One or more active compounds may be included in thecompositions of the invention, which may be prepared by any of thewell-known techniques of pharmacy comprising admixing the components,optionally including one or more accessory ingredients.

In general, compositions may be prepared by uniformly and intimatelyadmixing the active compound with a liquid or finely divided solidcarrier, or both, and then, if necessary, shaping the resulting mixture.For example, a tablet may be prepared by compressing or molding a powderor granules containing the active compound, optionally with one or moreaccessory ingredients. Compressed tablets may be prepared bycompressing, in a suitable machine, the compound in a free-flowing form,such as a powder or granules optionally mixed with a binder, lubricant,inert diluent, and/or surface active/dispersing agent(s). Molded tabletsmay be made by molding, in a suitable machine, the powdered compoundmoistened with an inert liquid binder.

The compositions of the invention include those suitable for oral,rectal, topical, buccal (e.g., sub-lingual), vaginal, parenteral (e.g.,subcutaneous, intramuscular, intradermal, or intravenous), topical(i.e., both skin and mucosal surfaces, including airway surfaces) andtransdermal administration, although the most suitable route in anygiven case will depend on the nature and severity of the condition beingtreated and on the nature of the particular active compound that isbeing used. Preferred routes of parenteral administration includeintrathecal injection and intraventricular injection into a ventricle ofthe brain.

Compositions suitable for oral administration may be presented indiscrete units, such as capsules, cachets, lozenges, or tablets, eachcontaining a predetermined amount of the active compound; as a powder orgranules; as a solution or a suspension in an aqueous or non-aqueousliquid; or as an oil-in-water or water-in-oil emulsion. Suchcompositions may be prepared by any suitable method of pharmacy, whichincludes the step of bringing into association the active compound and asuitable carrier (which may contain one or more accessory ingredients asnoted above).

Compositions suitable for buccal (sub-lingual) administration includelozenges comprising the active compound in a flavored base, usuallysucrose and acacia or tragacanth; and pastilles comprising the compoundin an inert base such as gelatin and glycerin or sucrose and acacia.

Compositions of the present invention suitable for parenteraladministration comprise sterile aqueous and non-aqueous injectionsolutions of the active compound, which preparations are preferablyisotonic with the blood of the intended recipient. These preparationsmay contain anti-oxidants, buffers, bacteriostats and solutes thatrender the composition isotonic with the blood of the intendedrecipient. Aqueous and non-aqueous sterile suspensions may includesuspending agents and thickening agents. The compositions may bepresented in unit/dose or multi-dose containers, for example sealedampoules and vials, and may be stored in a freeze-dried (lyophilized)condition requiring only the addition of the sterile liquid carrier, forexample, saline or water-for-injection immediately prior to use.Extemporaneous injection solutions and suspensions may be prepared fromsterile powders, granules and tablets of the kind previously described.

For example, in one aspect of the present invention, there is providedan injectable, stable, sterile composition comprising a compound ofFormula (I), Formula (II), Formula (III), Formula (IV), Formula (V), ora salt or prodrug thereof, in a unit dosage form in a sealed container.The compound or salt is provided in the form of a lyophilizate that iscapable of being reconstituted with a suitable pharmaceuticallyacceptable carrier to form a liquid composition suitable for injectionthereof into a subject. The unit dosage form typically comprises fromabout 10 mg to about 10 grams of the compound or salt. When the compoundor salt is substantially water-insoluble, a sufficient amount ofemulsifying agent that is physiologically acceptable may be employed insufficient quantity to emulsify the compound or salt in an aqueouscarrier. One such useful emulsifying agent is phosphatidyl choline.

Compositions suitable for rectal administration are preferably presentedas unit dose suppositories. These may be prepared by mixing the activecompound with one or more conventional solid carriers, for example,cocoa butter, and then shaping the resulting mixture.

Compositions suitable for topical application to the skin preferablytake the form of an ointment, cream, lotion, paste, gel, spray, aerosol,or oil. Carriers that may be used include petroleum jelly, lanoline,polyethylene glycols, alcohols, transdermal enhancers, and combinationsof two or more thereof.

Compositions suitable for transdermal administration may be presented asdiscrete patches adapted to remain in intimate contact with theepidermis of the recipient for a prolonged period of time. Compositionssuitable for transdermal administration may also be delivered byiontophoresis (see, for example, Pharmaceutical Research 3 (6):318(1986)) and typically take the form of an optionally buffered aqueoussolution of the active compound.

Also provided in some embodiments are compositions comprising an activecompound and a biocide. A “biocide” as used herein refers to a substancewith the ability to kill or to inhibit the growth of microorganisms(e.g., bacteria, fungal cells, protozoa, etc.), which substance is notan active compound give above in Section B. Common biocides includeoxidizing and non-oxidizing chemicals. Examples of oxidizing biocidesinclude chlorine, chlorine dioxide, and ozone. Examples of non-oxidizingbiocides include quaternary ammonium compounds, formaldehyde, andanionic and non-anionic surface agents. Chlorine is the most commonbiocide used in sanitizing water systems.

An “antibiotic” as used herein is a type of “biocide.” Commonantibiotics include aminoglycosides, carbacephems (e.g., loracarbef),carbapenems, cephalosporins, glycopeptides (e.g., teicoplanin andvancomycin), macrolides, monobactams (e.g., aztreonam) penicillins,polypeptides (e.g., bacitracin, colistin, polymyxin B), quinolones,sulfonamides, tetracyclines, etc. Antibiotics treat infections by eitherkilling or preventing the growth of microorganisms. Many act to inhibitcell wall synthesis or other vital protein synthesis of themicroorganisms.

Aminoglycosides are commonly used to treat infections caused byGram-negative bacteria such as Escherichia coli and Klebsiella,particularly Pseudomonas aeroginosa. Examples of aminoglycosidesinclude, but are not limited to amikacin, gentamicin, kanamycin,neomycin, netilmicin, streptomycin, tobramycin, and paromomycin.

Carbapenems are broad-spectrum antibiotics, and include, but are notlimited to, ertapenem, doripenem, imipenem/cilstatin, and meropenem.

Cephalosporins include, but are not limited to, cefadroxil, cefazolin,cefalotin (cefalothin), cefalexin, cefaclor, cefamandole, cefoxitin,cefprozil, loracarbef, cefuroxime, cefixime, cefdinir, cefditoren,cefoperazone, cefotaxime, cefpodoxime, ceftazidime, ceftibuten,ceftizoxime, ceftriaxone, cefepime, cefpirome, and ceftobiprole.

Macrolides include, but are not limited to, azithromycin,clarithromycin, dirithromycin, erythromycin, roxithromycin,troleandomycin, telithromycin and spectinomycin.

Penicillins include, but are not limited to, amoxicillin, ampicillin,azlocillin, bacampicillin, carbenicillin, cloxacillin, dicloxacillin,flucloxacillin, mezlocillin, meticillin, nafcillin, oxacillin,penicillin, piperacillin and ticarcillin.

Quinolones include, but are not limited to, ciprofloxacin, enoxacin,gatifloxacin, gemifloxacin, levofloxacin, lomefloxacin, moxifloxacin,norfloxacin, ofloxacin and trovafloxacin.

Sulfonamides include, but are not limited to, mafenide, prontosil,sulfacetamide, sulfamethizole, sulfanilamide, sulfasalazine,sulfisoxazole, trimethoprim, and co-trimoxazole(trimethoprim-sulfamethoxazole).

Tetracyclines include, but are not limited to, demeclocycline,doxycycline, minocycline, oxytetracycline and tetracycline.

Other antibiotics include arsphenamine, chloramphenicol, clindamycin,lincomycin, ethambutol, fosfomycin, fusidic acid, furazolidone,isoniazid, linezolid, metronidazole, mupirocin, nitrofurantoin,platensimycin, pyrazinamide, quinupristin/dalfopristin, rifampin(rifampicin), tinidazole, etc.

In some embodiments, a dentifrice composition is provided comprising theactive compounds. A “dentifrice” is a substance that is used to cleanthe teeth. It may be in the form of, e.g., a paste or powder. Commonlyknown dentifrices include toothpaste, mouthwash, chewing gum, dentalfloss, and dental cream. Other examples of dentifrices includetoothpowder, mouth detergent, troches, dental or gingival massage cream,dental strips, dental gels, and gargle tablets. Examples of dentifricecompositions comprising toothpaste and mouthwash are found in U.S. Pat.No. 6,861,048 (Yu et al.); U.S. Pat. No. 6,231,836 (Takhtalian et al.);and U.S. Pat. No. 6,331,291 (Glace et al.); each incorporated byreference herein in their entirety.

A coating composition is also provided. A “coating” as used herein isgenerally known. Any of a variety of organic and aqueous coatingcompositions, with or without pigments, may be modified to containbiofilm inhibiting compositions as described herein, including but notlimited to those described in U.S. Pat. Nos. 7,109,262, 6,964,989,6,835,459, 6,677,035, 6,528,580, 6,235,812, etc., each incorporated byreference herein in their entirety.

In general, the coatings comprise a film-forming resin, an aqueous ororganic solvent that disperses the resin; and, optionally, at least onepigment. Other ingredients such as colorants, secondary pigments,stabilizers and the like can be included if desired. However, for use inthe present invention the compositions further comprise one or morebiofilm inhibiting compounds as described herein, which may be carriedby or dispersed in the solvent and/or resin, so that the biofilminhibiting compounds are dispersed or distributed on the substrate anarticle coated. A resin may carry the biofilm inhibiting compoundsthrough covalent attachment through means well known in the art. Theresin may comprise, for example, a polymeric material. A polymericmaterial is a material that is comprised of large molecules made fromassociated smaller repeating structural units, often covalently linked.Common examples of polymeric materials are unsaturated polyester resins,and epoxy resins.

Any suitable article can be coated, in whole or in part, with acomposition of the invention. Suitable articles include, but are notlimited to, automobiles and airplanes (including substrates such as wingand propeller surfaces for aerodynamic testing), vessel hulls (includinginterior and exterior surfaces thereof), pressure vessels (includinginterior and exterior surfaces thereof) medical implants, windmills,etc. Coating of the article with the composition can be carried out byany suitable means, such as by brushing, spraying, electrostaticdeposition, dip coating, doctor blading, etc.

D. Methods of Use

Methods of controlling biofilm formation on a substrate are disclosed,comprising the step of administering an active compound to a substratein an amount effective to inhibit biofilm formation. A “substrate” asused herein is a base on which an organism, such as those commonly foundin biofilms, may live. The term “substrate,” as used herein, refers toany substrate, whether in an industrial or a medical setting, thatprovides or can provide an interface between an object and a fluid,permitting at least intermittent contact between the object and thefluid. A substrate, as understood herein, further provides a plane whosemechanical structure, without further treatment, is compatible with theadherence of microorganisms. Substrates compatible with biofilmformation may be natural or synthetic, and may be smooth or irregular.Fluids contacting the substrates can be stagnant or flowing, and canflow intermittently or continuously, with laminar or turbulent or mixedflows. A substrate upon which a biofilm forms can be dry at times withsporadic fluid contact, or can have any degree of fluid exposureincluding total immersion. Fluid contact with the substrate can takeplace via aerosols or other means for air-borne fluid transmission.

Biofilm formation with health implications can involve those substratesin all health-related environments, including substrates found inmedical environments and those substrates in industrial or residentialenvironments that are involved in those functions essential to humanwell being, for example, nutrition, sanitation and the prevention ofdisease. Substrates found in medical environments include the inner andouter aspects of various instruments and devices, whether disposable orintended for repeated uses. Examples include the entire spectrum ofarticles adapted for medical use, including scalpels, needles, scissorsand other devices used in invasive surgical, therapeutic or diagnosticprocedures; implantable medical devices, including artificial bloodvessels, catheters and other devices for the removal or delivery offluids to patients, artificial hearts, artificial kidneys, orthopedicpins, plates and implants; catheters and other tubes (includingurological and biliary tubes, endotracheal tubes, peripherablyinsertable central venous catheters, dialysis catheters, long termtunneled central venous catheters, peripheral venous catheters, shortterm central venous catheters, arterial catheters, pulmonary catheters,Swan-Ganz catheters, urinary catheters, peritoneal catheters), urinarydevices (including long term urinary devices, tissue bonding urinarydevices, artificial urinary sphincters, urinary dilators), shunts(including ventricular or arterio-venous shunts); prostheses (includingbreast implants, penile prostheses, vascular grafting prostheses, heartvalves, artificial joints, artificial larynxes, otological implants),vascular catheter ports, wound drain tubes, hydrocephalus shunts,pacemakers and implantable defibrillators, and the like. Other exampleswill be readily apparent to practitioners in these arts. Substratesfound in the medical environment also include the inner and outeraspects of pieces of medical equipment, medical gear worn or carried bypersonnel in the health care setting. Such substrates can includecounter tops and fixtures in areas used for medical procedures or forpreparing medical apparatus, tubes and canisters used in respiratorytreatments, including the administration of oxygen, of solubilized drugsin nebulizers and of anesthetic agents. Also included are thosesubstrates intended as biological barriers to infectious organisms inmedical settings, such as gloves, aprons and faceshields. Commonly usedmaterials for biological barriers may be latex-based or non-latex based.Vinyl is commonly used as a material for non-latex surgical gloves.Other such substrates can include handles and cables for medical ordental equipment not intended to be sterile. Additionally, suchsubstrates can include those non-sterile external substrates of tubesand other apparatus found in areas where blood or body fluids or otherhazardous biomaterials are commonly encountered.

Substrates in contact with liquids are particularly prone to biofilmformation. As an example, those reservoirs and tubes used for deliveringhumidified oxygen to patients can bear biofilms inhabited by infectiousagents. Dental unit waterlines similarly can bear biofilms on theirsubstrates, providing a reservoir for continuing contamination of thesystem of flowing an aerosolized water used in dentistry. Sprays,aerosols and nebulizers are highly effective in disseminating biofilmfragments to a potential host or to another environmental site. It isespecially important to health to prevent biofilm formation on thosesubstrates from where biofilm fragments can be carried away by sprays,aerosols or nebulizers contacting the substrate.

Other substrates related to health include the inner and outer aspectsof those articles involved in water purification, water storage andwater delivery, and articles involved in food processing. Substratesrelated to health can also include the inner and outer aspects of thosehousehold articles involved in providing for nutrition, sanitation ordisease prevention. Examples can include food processing equipment forhome use, materials for infant care, tampons and toilet bowls.“Substrate” as used herein also refers to a living substrate, such asthe inner ear of a patent.

Substrates can be smooth or porous, soft or hard. Substrates can includea drainpipe, glaze ceramic, porcelain, glass, metal, wood, chrome,plastic, vinyl, Formica® brand laminate, or any other material that mayregularly come in contact with an aqueous solution in which biofilms mayform and grow. The substrate can be a substrate commonly found onhousehold items such as shower curtains or liners, upholstery, laundry,and carpeting.

A substrate on which biofilm preventing, removing or inhibiting isimportant is that of a ship hull. Biofilms, such as those of Halomonaspacifica, promote the corrosion of the hull of ships and also increasethe roughness of the hull, increasing the drag on the ship and therebyincreasing fuel costs. The biofilm can also promote the attachment oflarger living structures such as barnacles on the ship hull. Fuel canaccount for half of the cost of marine shipping, and the loss in fuelefficiency due to biofilm formation is substantial.

Substrates on which biofilms can adhere include those of livingorganisms, as in the case of humans with chronic infections caused bybiofilms, as discussed above. Biofilms can also form on the substratesof food contact surfaces, such as those used for processing seafood, andalso on food products themselves. Examples of seafood products that mayhave biofilm contamination include oysters. Human infections caused bythe ingestion of raw oysters has been linked to Vibrio vulnificusbacterium. Vibrio bacteria attach to algae and plankton in the water andtransfer to the oysters and fish that feed on these organisms.

Other examples of substrates or devices on which biofilms can adhere canbe found in U.S. Pat. Nos. 5,814,668 and 7,087,661; and U.S. Pat.Application Publication Nos. 2006/0228384 and 2006/0018945, each ofwhich is incorporated herein by reference in its entirety.

In some embodiments, methods of enhancing the effects of a biocide aredisclosed, comprising the step of administering an active compound incombination with a biocide, the active compound being administered in anamount effective to enhance the effects of the biocide.

“Administering” or “administration of” an active compound and/or biocideas used herein in inclusive of contacting, applying, etc. (e.g.,contacting with an aqueous solution, contacting with a surface (e.g., ahospital surface such as a table, instrumentation, etc.)), in additionto providing to a subject (for example, to a human subject in need oftreatment for a microbial infection).

“Enhancing” the effects of a biocide by administering an active compoundin combination with the biocide refers to increasing the effectivenessof the biocide, such that the microorganism killing and/or growthinhibition is higher at a certain concentration of the biocideadministered in combination with the active compound than without. Insome embodiments, a bacteria or other microorganism is “sensitized” tothe effects of a biocide, such that the bacteria or other microorganismthat was resistant to the biocide prior to administering the activecompound (e.g., little to none, or less than 20, 10, 5 or 1% are killedupon application) is rendered vulnerable to that biocide upon or afteradministering the active compound (e.g., greater than 20, 30, 40, 50,60, 70, 80, 90, or 95% or more are killed).

As used herein, the administration of two or more compounds (inclusiveof active compounds and biocides) “in combination” means that the twocompounds are administered closely enough in time that theadministration of or presence of one alters the biological effects ofthe other. The two compounds may be administered simultaneously(concurrently) or sequentially.

Simultaneous administration of the compounds may be carried out bymixing the compounds prior to administration, or by administering thecompounds at the same point in time but at different anatomic sites orusing different routes of administration, or administered at timessufficiently close that the results observed are indistinguishable fromthose achieved when the compounds are administered at the same point intime.

Sequential administration of the compounds may be carried out byadministering, e.g., an active compound at some point in time prior toadministration of a biocide, such that the prior administration ofactive compound enhances the effects of the biocide (e.g., percentage ofmicroorganisms killed and/or slowing the growth of microorganisms). Insome embodiments, an active compound is administered at some point intime prior to the initial administration of a biocide. Alternatively,the biocide may be administered at some point in time prior to theadministration of an active compound, and optionally, administered againat some point in time after the administration of an active compound.

Also disclosed is a method of controlling biofilm formation wherein thebiofilm comprises Gram-negative bacteria. “Gram-negative” bacteria arethose that do not retain crystal violet dye after an alcohol wash in theGram staining protocol. This is due to structural properties in the cellwalls of the bacteria. Many genera and species of Gram-negative bacteriaare pathogenic. Gram-negative bacteria include members of the phylumproteobacteria, which include genus members Escherichia, Salmonella,Vibrio, and Helicobacter. A “genus” is a category of biologicalclassification ranking between the family and the species, comprisingstructurally or phylogenetically related species, or an isolated speciesexhibiting unusual differentiation. It is usually designated by a Latinor latinized capitalized singular noun. Examples of genera ofbiofilm-forming bacteria affected by active compounds of this inventioninclude, but are not limited to, Pseudomonas, Bordetella, Vibrio,Haemophilus, Halomonas, and Acinetobacter. “Species” refer to a categoryof biological classification ranking below the genus, and comprisemembers that are structurally or phylogenetically related, or anisolated member exhibiting unusual differentiation. Species are commonlydesignated by a two-part name, which name includes the capitalized anditalicized name of the genus in which the species belongs as the firstword in the name, followed by the second word that more specificallyidentifies the member of the genus, which is not capitalized. Examplesof species of bacteria capable of forming biofilms that are affected byactive compounds of the present invention include Pseudomonasaeuroginosa, Bordetella pertussis, Vibrio vulnificus, Haemophilusinfluenzae, Halomonas pacifica, and Acinetobacter baumannii.

Other examples of Gram-negative bacteria affected by active compounds ofthe present invention include, but are not limited to, bacteria of thegenera Klebsiella, Proteus, Neisseria, Helicobacter, Brucella,Legionella, Campylobacter, Francisella, Pasteurella, Yersinia,Bartonella, Bacteroides, Streptobacillus, Spirillum, Moraxella andShigella.

Examples of Gram-positive bacteria affected by active compounds of thepresent invention include, but are not limited to, bacteria of thegenera Listeria, Staphylococcus, Streptococcus, Bacillus,Corynebacterium, Peptostreptococcus, and Clostridium. Furthermore,bacteria affected by active compounds of the present invention includesGram-positive bacteria including, but not limited to, Listeriamonocytogenes, Staphylococcus aureus, Streptococcus pyogenes,Streptococcus pneumoniae, Bacillus cereus, Bacillus anthracis,Clostridium botulinum, Clostridium perfringens, Clostridium difficile,Clostridium tetani, Corynebacterium diphtheriae, Corynebacteriumulcerans, and Peptostreptococcus anaerobius. Additional bacteriaaffected by active compounds of the present invention also includebacterial genera including, but not limited to, Actinomyces,Propionibacterium, Nocardia and Streptomyces.

“Planktonic” bacteria are bacteria that are free-swimming in a fluid, asopposed to attached to a surface (such as in a biofilm).

Various nosocomial infections that are especially prevalent in intensivecare units implicate Acinetobacter species such as Acinetobacterbaumannii and Acinetobacter lwoffi. Acinetobacter baumanni is a frequentcause of nosocomial pneumonia, and can also cause skin and woundinfections and bacteremia. Acinetobacter lwoffi causes meningitis. TheAcinetobacter species are resistant to many classes of antibiotics. TheCDC has reported that bloodstream infections implicating Acinetobacterbaumanni were becoming more prevalent among service members injuredduring the military action in Iraq and Afghanistan.

Staphylococcus aureus is a common cause of nosocomial infections, oftenfound in post-surgical wound infections. Staphylococcus aureus can alsocause variety of other infections in humans (e.g., skin infections), aswell as contribute to mastitis in dairy cows. Methicillin-resistantStaphylococcus aureaus (MRSA), in particular, is especially difficult totreat due to multiple drug resistences, including penicillins andcephalosporins. MRSA has become problematic in hospital settings,particularly among the more susceptible patients with open wounds,invasice devices, weakened immune systems, etc.

A “fungal cell” as used herein may be any fungal cell belonging to thegenera including, but not limited to, Aspergillus, Candida,Cryptococcus, Coccidioides, Tinea, Sporothrix, Blastomyces, Histoplasma,Pneumocystis and Saccharomyces. Additionally, fungus includes, but isnot limited to, Aspergillus fumigatus, Aspergillus flavus, Aspergillusniger, Aspergillus terreus, Aspergillus nidulans, Candida albicans,Coccidioides immitis, Cryptococcus neoformans, Tinea unguium, Tineacorporis, Tinea cruris, Sporothrix schenckii, Blastomyces dermatitidis,Histoplasma capsulatum, Histoplasma duboisii, and Saccharomycescerevisiae.

“Protazoa” are unicellular eukaryotic microorganisms, and includeflagellates, amoeboids, sporozoans, ciliates, etc.

A method for treating a chronic bacterial infection in a subject in needthereof is disclosed, comprising administering active compound to saidsubject in an amount effective to inhibit, reduce, or remove a biofilmcomponent of said chronic bacterial infection. “Treating” as used hereinrefers to any type of activity that imparts a benefit to a patientafflicted with a disease, including improvement in the condition of thepatient (e.g., in one or more symptoms), delay in the progression of thedisease, delay in onset of the disease, etc. The present invention isprimarily concerned with the treatment of human subjects, but theinvention may also be carried out on animal subjects, particularlymammalian subjects (e.g., mice, rats, dogs, cats, rabbits, and horses),avian subjects (e.g., parrots, geese, quail, pheasant), livestock (e.g.,pigs, sheep, goats, cows, chickens, turkey, duck, ostrich, emu), reptileand amphibian subjects, for veterinary purposes or animal husbandry, andfor drug screening and drug development purposes.

A “chronic bacterial infection” is a bacterial infection that is of along duration or frequent recurrence. For example, a chronic middle earinfection, or otitis media, can occur when the Eustachian tube becomesblocked repeatedly due to allergies, multiple infections, ear trauma, orswelling of the adenoids. The definition of “long duration” will dependupon the particular infection. For example, in the case of a chronicmiddle ear infection, it may last for weeks to months. Other knownchronic bacterial infections include urinary tract infection (mostcommonly caused by Escherichia coli and/or Staphylococcussaprophyticus), gastritis (most commonly caused by Helicobacter pylori),respiratory infection (such as those commonly afflicting patents withcystic fibrosis, most commonly caused by Pseudomonas aeuroginosa),cystitis (most commonly caused by Escherichia coli), pyelonephritis(most commonly caused by Proteus species, Escherichia coli and/orPseudomonas species), osteomyelitis (most commonly caused byStaphylococcus aureus, but also by Escherichia coli), bacteremia, skininfection, rosacea, acne, chronic wound infection, infectious kidneystones (can be caused by Proteus mirabilis), bacterial endocarditis, andsinus infection. A common infection afflicting pigs is atrophic rhinitis(caused by Bordatella species, e.g. Bordatella rhinitis).

Also disclosed is a method of clearing a preformed biofilm from asubstrate comprising the step of administering an effective amount ofcompound to said substrate, wherein said effective amount will reducethe amount of said biofilm on said substrate. “Preformed biofilm” is abiofilm that has begun to adhere to a substrate. The biofilm may or maynot yet be fully formed.

E. Devices

Medical devices comprising a substrate and an effective amount of activecompound are also disclosed. “Medical device” as used herein refers toan object that is inserted or implanted in a subject or applied to asurface of a subject. Common examples of medical devices include stents,fasteners, ports, catheters, scaffolds and grafts. A “medical devicesubstrate” can be made of a variety of biocompatible materials,including, but not limited to, metals, ceramics, polymers, gels, andfluids not normally found within the human body. Examples of polymersuseful in fabricating medical devices include such polymers assilicones, rubbers, latex, plastics, polyanhydrides, polyesters,polyorthoesters, polyamides, polyacrylonitrile, polyurethanes,polyethylene, polytetrafluoroethylene, polyethylenetetraphthalate, etc.Medical devices can also be fabricated using naturally-occurringmaterials or treated with naturally-occurring materials. Medical devicescan include any combination of artificial materials, e.g., combinationsselected because of the particular characteristics of the components.Medical devices can be intended for short-term or long-term residencewhere they are positioned. A hip implant is intended for several decadesof use, for example. By contrast, a tissue expander may only be neededfor a few months, and is removed thereafter.

Some examples of medical devices are found in U.S. Pat. No. 7,081,133(Chinn et al.); U.S. Pat. No. 6,562,295 (Neuberger); and U.S. Pat. No.6,387,363 (Gruskin); each incorporated by reference herein in theirentirety.

F. Covalent Coupling of Active Compounds

Active compounds as described herein can be covalently coupled tosubstrates. Examples of substrates include solid supports and polymers.The polymers, typically organic polymers, may be in solid form, liquidform, dispersed or solubilized in a solvent (e.g., to form a coatingcomposition as described above), etc. The solid support may include thesubstrate examples as described above to be coated with or treated withactive compounds of the invention.

Covalent coupling can be carried out by any suitable technique. Activecompounds of the present invention may be appended to a substrate viaaldehyde condensation, amide or peptide bond, carbon-carbon bond, or anysuitable technique commonly used in the art.

Various coupling reactions can be used to covalently link activecompounds of the present invention to a substrate. Examples of couplingreactions that can be used include, but are not limited to, Hiyama,Suzuki, Sonogashira, Heck, Stille, Negishi, Kumada, Wurtz, Ullmann,Cadiot-Chodkiewicz, Buchwald-Hartwig, and Grignard reactions. Forexample, an active compound of Formula (I), Formula (II), Formula (III),Formula (IV) or Formula (V) that is substituted with a halo (e.g. bromoor chloro) can be coupled to a substrate via a Heck reaction.

This listing of examples of reactions that can be used to append activecompounds of the present invention to a substrate is not intended to beexhaustive. Those skilled in the art will readily appreciate variousother methods of carrying out these teachings. Further examples andexplanations of these types of reactions can be found in U.S. Pat. No.6,136,157 (Lindeberg et al.) and U.S. Pat. No. 7,115,653 (Baxter etal.), which are each hereby incorporated by reference in their entirety.

Some aspects of the present invention are described in more detail inthe following non-limiting examples.

Example 1: Synthesis of a 2-Aminoimidazole Library for Anti-BiofilmScreening Utilizing the Sonogashira Reaction

The synthesis of a 21 member library is described that employs ahigh-yielding Sonogashira reaction (Sonogashira et al., Tet. Lett. 1975,4467-4470; Chinchilla et al., Chem. Rev. 2007, 107, 874-922; Schnurch etal., European Journal of Organic Chemistry 2006, 3283-3307) on threearyl iodide protected 2-aminoimidazole scaffolds to generate smallmolecules for screening as anti-biofilm agents.

Access to the desired aryl halide intermediates for the Sonogashiracoupling was executed through the commercially available ortho, meta,and para substituted iodobenzoic acid derivatives 3-5 (Scheme 1).

Each was first transformed into its acid chloride before beingsequentially reacted with diazomethane and quenched with concentratedHBr. This afforded the requisite α-bromo ketones 6-8 in excellentyields. Installation of the 2-aminoimidazole subunit was achievedthrough condensation with Boc-guanidine (Zapf et al., Org. Lett. 2001,3, 1133-1136) in the presence of NaI. Attempts at performing theSonogashira reaction with intermediate 11 proved futile due to itsrelative insolubilty in all organic solvents except DMF. Low yields anddifficulty in purification of the desired product were observed,however, when DMF was used in the reaction. It therefore becamenecessary to protect the exocyclic amine functionality, and this wasaccomplished by reaction of derivatives 9-11 with LiHMDS in the presenceof Boc anhydride (Ando et al., Synlett 2006, 2836-2840). Despite theneed for the extra synthetic step, scaffolds 12-14 were quickly preparedon multi-gram scales in just three synthetic operations.

Attention then turned to a suitable catalyst system for the Sonogashirareaction. Lindel previously reported the use of PdCl₂(PPh₃)₂ as acatalyst for Heck and Sonogashira type reactions employing animidazole-based iminophosphorane intermediate (Poeverlein et al.,Synthesis-Stuttgart 2007, 3620-3626), and this provided the startingpoint for catalyst screening and reaction condition optimization (Scheme2).

SCHEME 2 Sonogashira Optimization

Entry Catalyst DIPEA 15 Solvent Yield (%)^(a) 1 Pd(PPh₃)₄ (5 mol %) 2equiv 3 equiv THF 48 2 PdCl₂(PPh₃)₂ (5 mol %) 2 equiv 3 equiv THF 71 3PdCl₂(PPh₃)₂ (5 mol %) 2 equiv 3 equiv Dioxane 65 4 PdCl₂(PPh₃)₂ (5 mol%) 2 equiv 3 equiv DMF 59 5 PdCl₂(PPh₃)₂ (5 mol %) 10 equiv 3 equiv THF78 6 PdCl₂(PPh₃)₂ (10 mol %) 10 equiv 5 equiv THF 90 ^(a)Isolated yield.

Conditions were screened using the p-iodo scaffold 14 and phenylacetylene 15 as the alkyne. PdCl₂(PPh₃)₂ was found to be a bettercatalyst at promoting the coupling than Pd(PPh₃)₄ in every conditionanalyzed. Increasing the catalyst load to 10 mol % was found to beoptimal, leading to complete consumption of the aryl iodide startingmaterial. This was essential because incomplete conversion led todifficulty in removing unreacted starting material which was laboriousand led to reduced reaction yields. THF was the best solvent among thethree scanned. Minor adjustments were also made in the amount of DIPEAused (10 equiv) and the alkyne coupling partner (5 equiv). With theconditions optimized on the trial system, the yield for the Sonogashiracoupling was 90% and was deemed acceptable for the synthesis of thelibrary.

Ten electronic and sterically diverse alkynes incorporating a number ofsynthetically relevant functionalities were then used to generate asmall library of 2-AI analogues using the optimized coupling conditions(Scheme 3).

SCHEME 3 Library Synthesis.

Entry Substrate Alkyne Yield (%) 1 12

94 2 13 (15) 87 3 12

97 4 13 (16) 91 5 14 (16) 94 6 12

81 7 13 (17) 90 8 14 (17) 80 9 12

92 10 14 (18) 85 Entry Substrate Alkyne Yield (%)^(a) 11 12

85 12 13 (19) 86 13 13

91 14 14

85 15 13

  81 ^(b) 16 14 (22)   80 ^(b) 17 13

86 18 14 (23) 90 19 14

82 20 14

84 ^(a)Isolated yield. ^(b) 8.0 equivalents of alkyne used.

Yields for all reactions were good to excellent (80-97%). The highestyielding reactions were those involving the 2,5-difluorophenyl alkynederivative 16 (Entries 3-5). TMS acetylene (Entry 14) and propargylalcohol (Entries 15 and 16) were also shown to be suitable couplingpartners under the extremely mild reaction conditions. Furthermore,consistently high yields were obtained when a single alkyne was coupledto all three regioisomers, indicating the robustness of the system.

In summary, the Sonogashira reaction was used to build a unique libraryof 21 complex 2-aminoimidazole based compounds that have the potentialto provide access to numerous other more advanced analogues. Thereaction proceeds in excellent yield regardless of the alkyne and arylhalide scaffold used. The biological screening of the analogues asmodulators of biofilm growth and maintenaince is currently underway andour findings will be reported in due course. Additionally, otherconditions may be utilized to take advantage of palladium catalyzedcross-couplings to create further libraries of 2-AI based molecules forevaluation as anti-biofilm agents.

Example 2: Synthesis of 2-Aminoimidazole Libraries for Anti-BiofilmScreening

Additional libraries are prepared as detailed in Scheme 4.

Example 3: Activity Testing of Compounds Against A. baumannii

Biofilm inhibition assays were performed by taking an overnight cultureof bacterial strain and subculturing it at an OD₆₀₀ of 0.10 into LBmedium. The compound being tested was then added at a predeterminedconcentration and then aliquoted (100 μL) into the wells of a 96-wellPVC microtiter plate (Wells not used for samples were filled with 100 μLof de-ionized water). Plates were then wrapped in GLAD Press n' Seal®and incubated under stationary conditions at 37° C. After 24 hours, themedia was discarded from the wells and the plates were washed thoroughlywith tap water. Plates were then stained with 100 μL of 0.1% solution ofcrystal violet (CV) and then incubated at an ambient temperature for 30minutes. Sample plates were then washed with tap water again, and theremaining stain was solubilized with 200 μL of 95% ethanol. Biofilminhibition was quantitated by measuring the OD₅₄₀ for each well bytransferring 125 μL of the solubilized CV stain into a polystyrenemicrotiter dish for analysis.

TABLE 1 Inhibition results from assays with A. Baumannii. COMPOUNDACTIVITY against A. baumannii

IC₅₀ < 50 μM

IC₅₀ = 6.54 μM

40 μM < IC₅₀ < 50 μM

IC₅₀ < 50 μM

50 μM < IC₅₀ < 75 μM

34% inhibition at 100 μM

Example 4: Synthesis of 2-Aminoimidazole Libraries for Anti-BiofilmScreening

Additional libraries were prepared having an amide on the2-aminoimidazole-phenyl structure.

Synthetic methods used for the library creation are outlined below inScheme 5.

The following compounds have been synthesized.

Detailed synthetic methods and characterizations are provided below formany of these compounds.

2-bromo-1-(2-iodophenyl)ethanone

2-Iodobenzoic acid (2.48 g, 10.0 mmol) was dissolved in anhydrousdichloromethane (30 mL) and cooled to 0° C. Oxalyl chloride (2.65 mL,30.0 mmol) was added drop-wise followed by the addition of a catalyticamount of anhydrous DMF (0.01 mL). The reaction was allowed to warm toroom temperature over the course of 1 h and after that time the solutionwas evaporated to dryness. The crude acid chloride was dissolved inanhydrous dichloromethane (10 mL) and added drop-wise to a 0° C.solution of CH₂N₂ (30.0 mmol generated from Diazald®/KOH) in diethylether (100 mL). This solution was stirred at 0° C. for 1 h upon whichthe reaction was quenched via the drop-wise addition of 48% solution ofconcentrated HBr (3.5 mL). The reaction mixture was diluted withdichloromethane (15 mL) and immediately washed with sat. NaHCO₃ (3×25mL) and brine (2×25 mL) before being dried (MgSO₄), filtered andconcentrated. The crude oil was purified via flash column chromatography(10-30% EtOAc/Hexanes) to obtain the desired compound2-bromo-1-(2-iodophenyl)ethanone (2.93 g, 90%) as a yellow oil: ¹H NMR(400 MHz, DMSO-d₆) δ 8.01 (d, 1H, J=7.6 Hz), 7.73 (d, 1H, J=8.0 Hz),7.53 (t, 1H, J=7.2 Hz), 7.27 (t, 1H, J=7.2 Hz), 4.85 (s, 2H); ¹³C NMR(100 MHz, DMSO-d₆) δ 195.0, 140.7, 140.5, 132.8, 129.0, 128.2, 93.2,36.1; HRMS (ESI) calcd for C₈H₇BrIO (MH⁺) 324.8719, found 324.8721.

2-bromo-1-(3-iodophenyl)ethanone

In a similar manner, 2.48 g (10.0 mmol) of 3-iodobenzoic acid afforded3.16 g (98%) of 2-bromo-1-(3-iodophenyl)ethanone as a colorless oil: ¹HNMR (400 MHz, DMSO-d₆) δ 8.29 (m, 1H), 8.02 (m, 2H), 7.35 (t, 1H, J=7.6Hz), 4.95 (s, 2H); ¹³C NMR (100 MHz, DMSO-d₆) δ 190.7, 142.2, 136.9,135.9, 131.0, 128.0, 95.3, 34.2; HRMS (ESI) calcd for C₈H₇BrIO (MH⁺)324.8719, found 324.8722.

2-bromo-1-(4-iodophenyl)ethanone

In a similar manner, 2.00 g (8.06 mmol) of 4-iodobenzoic acid afforded2.43 g (93%) of 2-bromo-1-(4-iodophenyl)ethanone as a yellow oil: ¹H NMR(300 MHz, DMSO-d₆) δ 7.94 (d, 2H, J=7.5 Hz), 7.72 (d, 2H, J=7.8 Hz),4.91 (s, 2H); ¹³C NMR (75 MHz, CDCl₃) δ 191.3, 138.8, 138.7, 133.7,131.5, 130.7, 102.8, 30.9; HRMS (ESI) calcd for C₈H₇BrIO (MH⁺) 324.8719,found 324.8718.

2-amino-5-(2-iodophenyl)imidazole-1-carboxylic acid tert-butyl ester

To a solution of 2-bromo-1-(2-iodophenyl)ethanone (2.70 g, 8.31 mmol) inanhydrous DMF (27 mL) was added Boc-guanidine (4.00 g, 24.9 mmol) andsodium iodide (2.47 g, 16.6 mmol). The reaction was stirred at ambienttemperature for 72 hours upon which the mixture was partitioned betweenEtOAc (150 mL) and water (75 mL). The organic layer was successivelywashed with water (3×50 mL) and brine (2×50 mL) before being dried(Na₂SO₄) and evaporated to dryness. The resulting crude oil was purifiedvia flash column chromatography (10-30% EtOAc/CH₂Cl₂) to obtain thetarget 2-amino-5-(2-iodophenyl)imidazole-1-carboxylic acid tert-butylester (1.84 g, 58%) as a tan foam: mp=121° C. (dec.); ¹H NMR (400 MHz,DMSO-d₆) δ 7.92 (m, 1H), 7.27 (dd, 1H, J=1.6, 8.0 Hz), 7.49 (s, 1H),7.41 (m, 1H), 7.00 (m, 1H), 6.59 (br s, 2H), 1.57 (s, 9H); ¹³C NMR (100MHz, DMSO-d₆) δ 149.6, 148.8, 140.3, 137.4, 130.1, 128.8, 128.1, 108.6,95.7, 85.0, 27.5; HRMS (ESI) calcd for C₁₄H₁₇IN₃O₂ (MO 386.0360, found386.0359.

2-amino-5-(3-iodophenyl)imidazole-1-carboxylic acid tert-butyl ester

In a similar manner, 2.50 g (7.71 mmol) of2-bromo-1-(3-iodophenyl)ethanone afforded 1.82 g (61%) of2-amino-5-(3-iodophenyl)imidazole-1-carboxylic acid tert-butyl ester asan off-white solid: mp=146-147° C.; ¹H NMR (400 MHz, DMSO-d₆) δ 8.11 (m,1H), 7.74 (m, 1H), 7.55 (m, 1H), 7.47 (s, 1H), 7.14 (m, 1H), 6.62 (br s,2H), 1.58 (s, 9H); ¹³C NMR (100 MHz, DMSO-d₆) δ 150.5, 148.9, 135.8,135.3, 133.0, 130.6, 123.9, 107.3, 95.0, 93.9, 84.8, 27.6; HRMS (ESI)calcd for C₁₄H₁₇IN₃O₂ (MH⁺) 386.0360, found 386.0361.

2-amino-5-(4-iodophenyl)imidazole-1-carboxylic acid tert-butyl ester

In a similar manner, 1.10 g (3.39 mmol) of2-bromo-1-(4-iodophenyl)ethanone afforded 1.43 g (55%) of2-amino-5-(4-iodophenyl)imidazole-1-carboxylic acid tert-butyl ester asan off-white solid: mp=140-141° C.; ¹H NMR (300 MHz, DMSO-d₆) δ 7.68 (d,2H, J=8.4 Hz), 7.53 (d, 2H, J=8.7 Hz), 7.42 (s, 1H), 6.61 (br s, 2H),1.57 (s, 9H); ¹³C NMR (100 MHz, DMSO-d₆) δ 150.5, 148.9, 137.1, 136.0,133.0, 126.8, 107.0, 106.9, 92.4, 84.8, 27.6; HRMS (ESI) calcd forC₁₄H₁₇IN₃O₂ (MH⁺) 386.0360, found 386.0358.

2-tert-butoxycarbonylamino-5-(2-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester

To a 0° C. solution of 2-amino-5-(2-iodophenyl)imidazole-1-carboxylicacid tert-butyl ester (1.01 g, 2.62 mmol) and di-tert-butyl dicarbonate(0.600 g, 2.75 mmol) in anhydrous THF (13 mL) was added drop-wise a 1Msolution of NaHMDS in THF (5.24 mL, 5.24 mmol). Upon completion thereaction was allowed to stir at 0° C. for an additional 15 minutesbefore the ice bath was removed and the reaction was permitted to warmto ambient temperature. It was then diluted with EtOAc (20 mL) andquenched with sat. NH₄Cl (10 mL). The aqueous layer was removed and theorganics were washed successively with sat. NaHCO₃ (2×20 mL) and brine(1×20 mL) before being dried (Na₂SO₄) and concentrated under reducedpressure. The crude oil was purified via flash column chromatography(10-40% EtOAc/Hexanes) to obtain the title compound2-tert-butoxycarbonylamino-5-(2-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester (0.937 g, 75%) as a white foam: mp=132-133° C.; ¹H NMR(400 MHz, DMSO-d₆) δ 9.60 (s, 1H), 7.96 (m, 1H), 7.89 (s, 1H), 7.70 (d,2H, J=8.0 Hz), 7.44 (m, 1H), 7.06 (m, 1H), 1.57 (s, 9H), 1.44 (s, 9H);¹³C NMR (100 MHz, DMSO-d₆) δ 153.1, 146.7, 140.3, 138.8, 137.8, 136.8,130.2, 129.3, 128.3, 115.0, 96.1, 85.5, 80.1, 28.0, 27.23; HRMS (ESI)calcd for C₁₉H₂₅IN₃O₄ (MH⁺) 486.0884, found 486.0892.

2-tert-butoxycarbonylamino-5-(3-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester

In a similar manner, 0.780 g (1.61 mmol) of2-amino-5-(3-iodophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.768 g (78%) of 2-tert-butoxycarbonylamino-5-(3-iodophenyl)imidazole-1-carboxylic acid tert-butyl ester as an off-white solid:mp=96-97° C.; ¹H NMR (400 MHz, DMSO-d₆) δ 9.57 (s, 1H), 8.18 (m, 1H),8.00 (s, 1H), 7.84 (d, 2H, J=7.6 Hz), 7.26 (m, 1H), 7.18 (t, 1H, J=7.6Hz), 1.57 (s, 9H), 1.44 (s, 9H); ¹³C NMR (100 MHz, DMSO-d₆) δ 152.8,146.7, 139.8, 135.8, 135.4, 134.9, 132.9, 130.8, 123.9, 113.6, 95.2,85.4, 80.1, 27.9, 27.3; HRMS (ESI) calcd for C₁₉H₂₄IN₃O₄Na (MNa⁺)508.0703, found 508.0703.

2-tert-butoxycarbonylamino-5-(4-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester

In a similar manner, 0.352 g (0.917 mmol) of2-amino-5-(4-iodophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.403 g (71%) of 2-tert-butoxycarbonylamino-5-(4-iodophenyl)imidazole-1-carboxylic acid tert-butyl ester as an off-white solid:mp=157° C. (dec.); ¹H NMR (300 MHz, DMSO-d₆) δ 9.56 (s, 1H), 7.95 (s,1H), 7.72 (m, 2H), 7.60 (m, 2H), 1.56 (s, 9H), 1.43 (s, 9H); ¹³C NMR (75MHz, DMSO-d₆) δ 152.9, 146.7, 139.7, 137.3, 136.1, 132.3, 126.8, 113.3,93.1, 85.4, 80.1, 27.9, 27.3; HRMS (ESI) calcd for C₁₉H₂₄IN₃O₄Na (MNa⁺)508.0703, found 508.0704.

2-tert-butoxycarbonylamino-5-(4-phenylethynylphenyl)imidazole-1-carboxylicacid tert-butyl ester

2-tert-butoxycarbonylamino-5-(4-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester (0.100 g, 0.206 mmol) was dissolved in anhydrous THF (8mL) and to this solution was added DIPEA (0.359 mL, 2.06 mmol), CuI(0.004 g, 0.021 mmol), and PdCl₂(PPh₃)₂ (0.014 g, 0.021 mmol). Thesolution was then degassed at ambient temperature for 10 minutes. Duringthis time a solution of the phenyl acetylene (0.105 g, 1.03 mmol) inanhydrous THF (3 mL) was also degassed. The solution of alkyne was addeddrop-wise to the solution of aryl iodide and the reaction was stirred atroom temperature for 12 h. The reaction was filtered through a Celite®pad and the filter cake rinsed with EtOAc (10 mL). The filtrate waswashed with sat. NH₄Cl (2×10 mL), brine (2×10 mL), and dried (Na₂SO₄).Filtration and evaporation afforded the crude product which was purifiedvia flash column chromatography (10-50% EtOAc/Hexanes) to obtain 0.085 g(90%) of the target 2-tert-butoxycarbonylamino-5-(4-phenylethynylphenyl)imidazole-1-carboxylic acid tert-butyl ester as a yellow solid:mp=131-132° C.; ¹H NMR (400 MHz, DMSO-d₆) δ 9.60 (s, 1H), 8.00 (s, 1H),7.86 (d, 2H, J=7.6 Hz), 7.42-7.57 (m, 7H), 1.58 (s, 9H), 1.44 (s, 9H);¹³C NMR (100 MHz, DMSO-d₆) δ 152.9, 146.8, 139.9, 136.3, 133.0, 131.7,131.4, 128.8, 124.9, 122.4, 126.8, 120.9, 113.7, 89.9, 89.5, 85.4, 80.1,27.9, 27.3; HRMS (ESI) calcd for C₂₇H₂₉N₃O₄Na (MNa⁺) 482.2050, found482.2051.

2-tert-butoxycarbonylamino-5-[4-(3,5-difluorophenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(4-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.096 g (94%) of2-tert-butoxycarbonylamino-5-[4-(3,5-difluorophenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a brown solid: mp=197° C. (dec.); ¹H NMR (300MHz, DMSO-d₆) δ 9.60 (s, 1H), 8.02 (s, 1H), 7.91 (d, 2H, J=8.7 Hz), 7.59(d, 2H, J=8.7 Hz), 7.34 (m, 3H), 1.58 (s, 9H), 1.44 (s, 9H); ¹³C NMR(100 MHz, DMSO-d₆) δ 163.5, 161.1, 152.8, 146.7, 139.9, 136.2, 133.9,134.6, 133.6, 131.9, 128.0, 125.2, 119.9, 114.8, 114.5, 114.0, 105.0,91.5, 87.6, 85.4, 80.1, 27.7, 27.3; HRMS (ESI) calcd for C₂₇H₂₇F₂N₃O₄Na(MNa⁺) 518.1861, found 518.1861.

2-tert-butoxycarbonylamino-5-[4-(2-methoxyphenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(4-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.080 g (80%) of2-tert-butoxycarbonylamino-5-[4-(2-methoxyphenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a yellow foam: mp=97-98° C.; ¹H NMR (400 MHz,DMSO-d₆) δ 9.60 (s, 1H), 7.99 (s, 1H), 7.85 (d, 2H, J=7.2 Hz), 7.53 (d,2H, J=7.6 Hz), 7.49 (m, 1H), 7.39 (t, 1H, J=8.0 Hz), 7.11 (d, 1H, J=8.4Hz), 6.97 (t, 1H, J=7.6 Hz), 3.86 (s, 3H), 1.58 (s, 9H), 1.43 (s, 9H);¹³C NMR (100 MHz, DMSO-d₆) δ 159.6, 152.9, 146.8, 139.9, 136.3, 133.1,132.7, 131.5, 130.4, 130.0, 124.9, 121.3, 120.5, 113.6, 111.4, 93.1,86.7, 85.4, 80.1, 55.7, 27.9, 27.3; HRMS (ESI) calcd for C₂₈H₃₂N₃O₅(MH⁺) 490.2336, found 490.2335.

2-tert-butoxycarbonylamino-5-[4-(4-methoxy-2-methylphenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(4-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.087 g (85%) of2-tert-butoxycarbonylamino-5-[4-(4-methoxy-2-methylphenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a yellow foam: mp=142-143° C.; ¹H NMR (400 MHz,DMSO-d₆) δ 9.59 (s, 1H), 7.98 (s, 1H), 7.86 (d, 2H, J=8.4 Hz), 7.53 (d,2H, J=8.4 Hz), 7.43 (d, 1H, J=8.4 Hz), 6.92 (d, 1H, J=2.4 Hz), 6.81 (dd,1H, J=2.4, 8.4 Hz), 3.78 (s, 3H), 2.45 (s, 3H), 1.57 (s, 9H), 1.44 (s,9H); ¹³C NMR (100 MHz, DMSO-d₆) δ 159.5, 152.9, 146.8, 141.5, 139.8,136.4, 133.0, 132.5, 131.4, 124.8, 121.6, 115.2, 114.3, 113.5, 111.8,92.0, 88.9, 85.4, 80.1, 55.2, 27.9, 27.3, 20.6; HRMS (ESI) calcd forC₂₉H₃₄N₃O₅ (MH⁺) 504.2492, found 504.2493.

2-tert-butoxycarbonylamino-5-(4-trimethylsilanylethynylphenyl)imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(4-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.080 g (85%) of2-tert-butoxycarbonylamino-5-(4-trimethylsilanylethynylphenyl)imidazole-1-carboxylicacid tert-butyl ester as a pale yellow solid: mp=88-89° C.; ¹H NMR (400MHz, DMSO-d₆) δ 9.59 (s, 1H), 7.99 (s, 1H), 7.81 (d, 2H, J=8.0 Hz), 7.46(d, 2H, J=8.4 Hz), 1.57 (s, 9H), 1.43 (s, 9H), 0.23 (s, 9H); ¹³C NMR (75MHz, DMSO-d₆) δ 152.8, 146.7, 139.9, 136.2, 133.2, 131.9, 124.8, 120.7,113.8, 105.3, 94.6, 85.4, 80.1, 27.9, 27.3, −0.07; HRMS (ESI) calcd forC₂₄H₃₃N₃O₄SiNa (MNa⁺) 478.2132, found 478.2135.

2-tert-butoxycarbonylamino-5-[4-(3-hydroxyprop-1-ynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(4-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.068 g (80%) of2-tert-butoxycarbonylamino-5-[4-(3-hydroxyprop-1-ynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a yellow solid: mp=76-77° C.; ¹H NMR (400 MHz,DMSO-d₆) δ 9.56 (s, 1H), 7.96 (s, 1H), 7.80 (d, 2H, J=7.2 Hz), 7.42 (d,2H, J=7.2 Hz), 5.36 (t, 1H, J=5.6 Hz), 4.31 (d, 2H, J=5.6 Hz), 1.57 (s,9H), 1.43 (s, 9H); ¹³C NMR (100 MHz, DMSO-d₆) δ 152.9, 146.8, 139.8,136.3, 132.7, 131.6, 124.8, 121.1, 113.6, 90.4, 85.4, 83.7, 80.1, 49.5,28.0, 27.3; HRMS (ESI) calcd for C₂₂H₂₇N₃O₅Na (MNa⁺) 436.1842, found436.1841.

2-tert-butoxycarbonylamino-5-[4-(3-dimethylaminoprop-1-ynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(4-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.081 g (90%) of2-tert-butoxycarbonylamino-5-[4-(3-dimethylaminoprop-1-ynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a yellow oil: ¹H NMR (400 MHz, DMSO-d₆) δ 9.58(s, 1H), 7.96 (s, 1H), 7.81 (d, 2H, J=8.8 Hz), 7.43 (d, 2H, J=8.4 Hz),3.46 (s, 2H), 2.25 (s, 6H), 1.57 (s, 9H), 1.44 (s, 9H); ¹³C NMR (75 MHz,DMSO-d₆) δ 152.9, 146.8, 139.8, 136.4, 132.5, 131.7, 124.8, 121.2,113.5, 85.8, 85.4, 85.0, 80.1, 47.8, 43.8, 27.9, 27.3; HRMS (ESI) calcdfor C₂₄H₃₂N₄O₄Na (MNa⁺) 463.2315, found 463.2313.

2-tert-butoxycarbonylamino-5-[4-(5-chloropent-1-ynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(4-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.062 g (82%) of2-tert-butoxycarbonylamino-5-[4-(5-chloropent-1-ynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a yellow solid: mp=61-62° C.; ¹H NMR (300 MHz,DMSO-d₆) δ 9.57 (s, 1H), 7.95 (s, 1H), 7.77 (m, 2H), 7.40 (m, 2H), 3.77(t, 2H, J=6.3 Hz), 2.59 (t, 2H, J=6.9 Hz), 1.99 (m, 2H), 1.57 (s, 9H),1.44 (s, 9H); ¹³C NMR (100 MHz, DMSO-d₆) δ 152.9, 146.8, 139.8, 137.3,136.4, 132.3, 131.6, 124.7, 121.6, 113.4, 89.4, 85.3, 81.2, 80.1, 44.4,31.1, 27.9, 27.3, 16.3; HRMS (ESI) calcd for C₂₄H₃₀ClN₃O₄Na (MNa⁺)482.1817, found 482.1816.

2-tert-butoxycarbonylamino-5-(4-oct-1-ynylphenyl)imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(4-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.081 g (84%) of2-tert-butoxycarbonylamino-5-(4-oct-1-ynylphenyl)imidazole-1-carboxylicacid tert-butyl ester as a yellow oil: ¹H NMR (400 MHz, CDCl₃) δ 9.17(s, 1H), 7.75 (d, 2H, J=8.0 Hz), 7.37 (d, 2H, J=8.0 Hz), 7.25 (s, 1H),2.40 (t, 2H, J=7.2 Hz), 1.63 (s, 9H), 1.60 (m, 2H), 1.54 (s, 9H), 1.45(m, 2H), 1.33 (m, 4H), 0.090 (t, 3H, J=6.4 Hz); ¹³C NMR (100 MHz,DMSO-d₆) δ 150.1, 149.5, 142.6, 138.6, 131.8, 127.0, 125.4, 123.3,108.1, 91.3, 86.9, 82.1, 80.9, 31.6, 29.0, 28.9, 28.5, 28.3, 28.2, 22.8,19.7, 14.3; HRMS (ESI) calcd for C₂₇H₃₈N₃O₄ (MH⁺) 468.2856, found468.2860.

2-tert-butoxycarbonylamino-5-(2-phenylethynylphenyl)imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(2-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.091 g (94%) of2-tert-butoxycarbonylamino-5-(2-phenylethynylphenyl)imidazole-1-carboxylicacid tert-butyl ester as a yellow solid: mp=113-114° C.; ¹H NMR (400MHz, DMSO-d₆) δ 9.62 (s, 1H), 8.20 (s, 1H), 8.02 (d, 1H, J=8.4 Hz), 7.62(m, 3H), 7.47 (m, 4H), 7.35 (m, 1H), 1.58 (s, 9H), 1.44 (s, 9H); ¹³C NMR(100 MHz, DMSO-d₆) δ 152.8, 146.7, 139.5, 134.9, 133.4, 133.4, 131.3,129.2, 129.1, 128.9, 127.3, 127.1, 122.1, 117.9, 115.1, 94.0, 89.5,85.5, 80.1, 27.9, 27.2; HRMS (ESI) calcd for C₂₇H₂₉N₃O₄Na (MNa⁺)482.2050, found 482.2050.

2-tert-butoxycarbonylamino-5-[2-(3,5-difluorophenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(2-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.099 g (97%) of2-tert-butoxycarbonylamino-5-[2-(3,5-difluorophenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a yellow foam: mp=161-162° C.; ¹H NMR (300 MHz,DMSO-d₆) δ 9.62 (s, 1H), 8.11 (s, 1H), 8.03 (m, 1H), 7.65 (m, 1H), 7.55(m, 1H), 7.34-7.44 (m, 4H), 1.49 (s, 9H), 1.44 (s, 9H); ¹³C NMR (75 MHz,DMSO-d₆) δ 164.1, 160.7, 152.7, 146.7, 139.6, 134.7, 133.9, 133.5,129.8, 127.3, 124.8, 117.1, 115.2, 114.8, 114.5, 105.3, 91.4, 91.4,85.5, 80.2, 27.9, 27.1; HRMS (ESI) calcd for C₂₇H₂₇F₂N₃O₄Na (MNa⁺)518.1861, found 518.1865.

2-tert-butoxycarbonylamino-5-[2-(2-methoxyphenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(2-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.081 g (81%) of2-tert-butoxycarbonylamino-5-[2-(2-methoxyphenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a tan foam: mp=134-135° C.; ¹H NMR (300 MHz,DMSO-d₆) δ 9.62 (s, 1H), 8.33 (s, 1H), 8.06 (d, 1H, J=7.2 Hz), 7.43-7.60(m, 4H), 7.33 (t, 1H, J=6.3 Hz), 7.13 (d, 1H, J=8.4 Hz), 7.02 (t, 1H,J=7.8 Hz), 3.87 (s, 3H), 1.46 (s, 9H), 1.44 (s, 9H); ¹³C NMR (100 MHz,DMSO-d₆) δ 159.8, 153.0, 146.7, 139.2, 134.9, 133.4, 133.1, 133.0,130.7, 128.9, 127.2, 126.9, 120.6, 118.3, 115.4, 111.4, 111.1, 92.9,91.2, 85.4, 80.1, 55.8, 27.9, 27.2; HRMS (ESI) calcd for C₂₈H₃₂N₃O₅(MH⁺) 490.2336, found 490.2332.

2-tert-butoxycarbonylamino-5-[2-(4-methoxy-2-methylphenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(2-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.095 g (92%) of2-tert-butoxycarbonylamino-5-[2-(4-methoxy-2-methylphenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a yellow foam: mp=59-60° C.; ¹H NMR (300 MHz,DMSO-d₆) δ 9.60 (s, 1H), 8.21 (s, 1H), 8.00 (d, 1H, J=7.5 Hz), 7.61 (d,1H, J=7.8 Hz), 7.47 (m, 2H), 7.33 (t, 1H, J=6.9 Hz), 6.96 (d, 1H, J=2.1Hz), 6.81 (dd, 1H, J=2.7, 8.7 Hz), 3.78 (s, 3H), 2.46 (s, 3H), 1.47 (s,9H), 1.44 (s, 9H); ¹³C NMR (75 MHz, DMSO-d₆) δ 159.7, 152.8, 146.7,141.6, 139.4, 135.1, 133.4, 133.1, 132.9, 128.7, 127.3, 127.0, 118.7,115.3, 115.0, 114.1, 111.9, 93.2, 91.8, 85.5, 80.1, 55.3, 27.9, 27.2,20.7; HRMS (ESI) calcd for C₂₉H₃₄N₃O₅ (MH⁺) 504.2492, found 504.2495.

2-tert-butoxycarbonylamino-5-[2-(3-chlorophenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(2-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.086 g (85%) of2-tert-butoxycarbonylamino-5-[2-(3-chlorophenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as an orange foam: mp=61-62° C.; ¹H NMR (300 MHz,DMSO-d₆) δ 9.61 (s, 1H), 8.15 (s, 1H), 8.04 (m, 1H), 8.01-8.14 (m, 2H),7.48-7.69 (m, 4H), 7.36 (m, 1H), 1.48 (s, 9H), 1.44 (s, 9H); ¹³C NMR (75MHz, DMSO-d₆) δ 152.8, 146.6, 139.5, 134.8, 133.6, 133.5, 130.7, 130.7,130.1, 129.5, 129.2, 127.3, 127.1, 124.1, 117.5, 115.2, 92.3, 90.7,85.5, 80.2, 27.9, 27.2; HRMS (ESI) calcd for C₂₇H₂₈ClN₃O₄Na (MNa⁺)516.1660, found 516.1658.

2-tert-butoxycarbonylamino-5-(3-phenylethynylphenyl)imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(3-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.084 g (87%) of2-tert-butoxycarbonylamino-5-(3-phenylethynylphenyl)imidazole-1-carboxylicacid tert-butyl ester as an orange foam: mp=76-77° C.; ¹H NMR (300 MHz,DMSO-d₆) δ 9.58 (s, 1H), 8.04 (s, 1H), 8.01 (m, 1H), 7.85 (m, 1H), 7.59(m, 2H), 7.44 (m, 5H), 1.58 (s, 9H), 1.44 (s, 9H); ¹³C NMR (100 MHz,DMSO-d₆) δ 152.9, 146.8, 139.8, 136.1, 133.2, 131.4, 130.1, 129.2,128.8, 127.4, 125.0, 122.6, 122.3, 113.5, 115.1, 89.4, 89.3, 85.4, 80.1,27.9, 27.3; HRMS (ESI) calcd for C₂₇H₂₉N₃O₄Na (MNa⁺) 482.2050, found482.2053.

2-tert-butoxycarbonylamino-5-[3-(3,5-difluorophenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(3-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.093 g (91%) of2-tert-butoxycarbonylamino-5-[3-(3,5-difluorophenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a tan solid: mp=69° C. (dec.); ¹H NMR (300 MHz,DMSO-d₆) δ 9.58 (s, 1H), 8.04 (s, 1H), 8.02 (s, 1H), 7.90 (m, 1H), 7.48(m, 1H), 7.37 (m, 3H), 1.58 (s, 9H), 1.44 (s, 9H); ¹³C NMR (100 MHz,DMSO-d₆) δ 163.6, 161.2, 152.8, 146.8, 139.8, 136.0, 133.3, 130.3,129.3, 127.7, 125.6, 125.2, 121.7, 114.9, 114.6, 113.5, 105.2, 91.2,87.1, 85.4, 80.1, 27.9, 27.3; HRMS (ESI) calcd for C₂₇H₂₇F₂N₃O₄Na (MNa⁺)518.1861, found 518.1866.

2-tert-butoxycarbonylamino-5-[3-(2-methoxyphenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(3-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.090 g (90%) of2-tert-butoxycarbonylamino-5-[3-(2-methoxyphenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as an orange foam: mp=75-76° C.; ¹H NMR (300 MHz,DMSO-d₆) δ 9.58 (s, 1H), 8.04 (s, 1H), 7.96 (s, 1H), 7.85 (m, 1H), 7.52(m, 1H), 7.41 (m, 3H), 7.11 (d, 1H, J=8.1 Hz), 6.98 (t, 1H, J=7.5 Hz),3.87 (s, 3H), 1.58 (s, 9H), 1.44 (s, 9H); ¹³C NMR (75 MHz, DMSO-d₆) δ159.7, 152.8, 146.8, 139.8, 136.1, 133.6, 130.5, 130.0, 129.1, 127.2,124.8, 123.1, 120.5, 113.5, 111.4, 111.2, 86.2, 85.3, 80.1, 55.7, 27.9,27.3; HRMS (ESI) calcd for C₂₈H₃₂N₃O₅ (MH⁺) 490.2336, found 490.2333.

2-tert-butoxycarbonylamino-5-[3-(3-chlorophenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(3-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.087 g (86%) of2-tert-butoxycarbonylamino-5-[3-(3-chlorophenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as an off-white foam: mp=67-68° C.; ¹H NMR (300MHz, DMSO-d₆) δ 9.58 (s, 1H), 8.03 (m, 2H), 7.86 (m, 1H), 7.66 (m, 1H),7.45-7.57 (m, 5H), 7.36 (m, 1H), 1.48 (s, 9H), 1.44 (s, 9H); ¹³C NMR(100 MHz, DMSO-d₆) δ 152.8, 146.8, 139.8, 136.0, 133.4, 133.2, 130.9,130.7, 130.2, 130.1, 129.2, 129.0, 127.6, 125.3, 124.2, 122.1, 113.5,90.5, 87.8, 85.4, 80.1, 27.9, 27.3; HRMS (ESI) calcd for C₂₇H₂₈ClN₃O₄Na(MNa⁺) 516.1660, found 516.1659.

2-tert-butoxycarbonylamino-5-[3-(4-pentylphenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(3-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.099 g (91%) of2-tert-butoxycarbonylamino-5-[3-(4-pentylphenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a white foam: mp=51-52° C.; ¹H NMR (300 MHz,DMSO-d₆) δ 9.58 (s, 1H), 8.03 (s, 1H), 7.98 (m, 1H), 7.82 (m, 1H),7.40-7.50 (m, 4H), 7.25 (d, 2H, J=8.4 Hz), 2.60 (t, 2H, J=7.5 Hz), 1.59(m, 2H), 1.57 (s, 9H), 1.44 (s, 9H), 1.24 (m, 4H), 0.861 (t, 3H, J=6.9Hz); ¹³C NMR (100 MHz, DMSO-d₆) δ 152.8, 146.8, 143.5, 139.8, 136.1,133.2, 131.3, 130.0, 129.1, 127.4, 124.8, 122.8, 119.5, 113.4, 89.6,88.7, 85.3, 80.1, 35.0, 30.9, 30.4, 27.9, 27.3, 21.9, 13.9, HRMS (ESI)calcd for C₃₂H₃₉N₃O₄Na (MNa⁺) 552.2832, found 552.2829.

2-tert-butoxycarbonylamino-5-[3-(3-hydroxyprop-1-ynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(3-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.069 g (81%) of2-tert-butoxycarbonylamino-5-[3-(3-hydroxyprop-1-ynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a white foam: mp=77-78° C.; ¹H NMR (300 MHz,DMSO-d₆) δ 9.56 (s, 1H), 8.01 (s, 1H), 7.88 (m, 1H), 7.93 (m, 1H), 7.39(t, 1H, J=7.8 Hz), 7.31 (m, 1H), 5.36 (t, 1H, J=5.7 Hz), 4.31 (d, 2H,J=6.0 Hz), 1.57 (s, 9H), 1.43 (s, 9H); ¹³C NMR (75 MHz, DMSO-d₆) δ152.8, 146.8, 139.8, 136.1, 133.1, 129.9, 129.0, 127.4, 124.7, 122.8,113.4, 89.9, 85.3, 83.5, 80.1, 49.1, 27.9, 27.3; HRMS (ESI) calcd forC₂₂H₂₇N₃O₅Na (MNa⁺) 436.1842, found 436.1838.

2-tert-butoxycarbonylamino-5-[3-(3-dimethylaminoprop-1-ynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.206 mmol) of2-tert-butoxycarbonylamino-5-(3-iodophenyl)imidazole-1-carboxylic acidtert-butyl ester afforded 0.077 g (86%) of2-tert-butoxycarbonylamino-5-[3-(3-dimethylaminoprop-1-ynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a white foam: mp=68-69° C.; ¹H NMR (300 MHz,DMSO-d₆) δ 9.57 (s, 1H), 8.00 (s, 1H), 7.87 (m, 1H), 7.79 (m, 1H), 7.38(t, 1H, J=7.5 Hz), 7.33 (m, 1H), 3.47 (s, 2H), 2.26 (s, 6H), 1.57 (s,9H), 1.43 (s, 9H); ¹³C NMR (100 MHz, DMSO-d₆) δ 152.9, 146.8, 139.7,136.1, 133.1, 132.0, 131.5, 131.4, 130.1, 129.0, 128.8, 128.7, 127.4,124.6, 122.9, 113.4, 85.4, 85.3, 84.9, 80.1, 47.7, 43.8, 27.9, 27.3;HRMS (ESI) calcd for C₂₄H₃₂N₄O₄Na (MNa⁺) 463.2315, found 463.2317.

5-(4-phenylethynylphenyl)-1H-imidazol-2-ylamine hydrochloride

2-tert-butoxycarbonylamino-5-(4-phenylethynylphenyl)imidazole-1-carboxylicacid tert-butyl ester (0.039 g, 0.109 mmol) was dissolved in anhydrousdichloromethane (5 mL) and cooled to 0° C. Trifluoroacetic acid (0.250mL) was added drop-wise while the reaction continued to stir at 0° C.Upon completion, the reaction was allowed to warm to room temperatureover the course of 12 h. Toluene (2 mL) was added and the reaction wasevaporated to dryness. The crude TFA salt was then dissolved indichloromethane (3 mL) and a 2M solution of HCl in diethyl ether (0.10mL) was added. The solution was again concentrated under reducedpressure and the resulting product triturated with cold diethyl ether (5mL) to afford 0.031 g (97%) of5-(4-phenylethynylphenyl)-1H-imidazol-2-ylamine in its correspondinghydrochloride salt form as a brown solid: ¹H NMR (300 MHz, DMSO-d₆) δ13.06 (s, 1H), 12.27 (s, 1H), 7.74 (d, 2H, J=8.7 Hz), 7.62 (d, 2H, J=8.7Hz), 7.55 (m, 5H), 7.43 (m, 3H); ¹³C NMR (75 MHz, DMSO-d₆) δ 147.9,131.9, 131.2, 128.9, 128.7, 127.9, 125.7, 124.3, 122.1, 121.4, 110.6,90.3, 89.0; HRMS (ESI) calcd for C₁₇H₁₃N₃ (M⁺) 259.1110, found 259.1113.

5-[4-(3,5-difluorophenylethynyl)phenyl]-1H-imidazol-2-ylaminehydrochloride

In a similar manner, 0.036 g (0.072 mmol) of2-tert-butoxycarbonylamino-5-[4-(3,5-difluorophenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester afforded 0.020 g (83%) of5-[4-(3,5-difluorophenylethynyl)phenyl]-1H-imidazol-2-ylamine in itscorresponding hydrochloride salt form as a pale yellow solid: ¹H NMR(400 MHz, DMSO-d₆) δ 13.10 (s, 1H), 12.29 (s, 1H), 7.76 (d, 2H, J=8.4Hz), 7.65 (d, 2H, J=8.4 Hz), 7.55 (m, 3H), 7.35 (m, 3H); ¹³C NMR (75MHz, DMSO-d₆) δ 163.8, 160.6, 147.9, 132.1, 128.6, 125.2, 124.3, 120.4,114.5, 110.9, 105.2, 91.0, 88.0; HRMS (ESI) calcd for C₁₇H₁₁F₂N₃ (M⁺)295.0921, found 295.0925.

5-(2-phenylethynylphenyl)-1H-imidazol-2-ylamine hydrochloride)

In a similar manner, 0.046 g (0.127 mmol) of2-tert-butoxycarbonylamino-5-(2-phenylethynylphenyl)imidazole-1-carboxylicacid tert-butyl ester afforded 0.037 g (97%) of5-(2-phenylethynylphenyl)-1H-imidazol-2-ylamine in its correspondinghydrochloride salt form as a brown amorphous solid: ¹H NMR (300 MHz,DMSO-d₆) δ 12.87 (s, 1H), 12.12 (s, 1H), 7.51-7.73 (m, 8H), 7.46 (m,4H); ¹³C NMR (75 MHz, DMSO-d₆) δ 147.3, 133.5, 131.3, 129.2, 129.1,128.6, 128.2, 126.9, 124.3, 121.9, 118.9, 112.2, 94.0, 88.2; HRMS (ESI)calcd for C₁₇H₁₃N₃ (M⁺) 259.1110, found 259.1110.

5-[2-(4-methoxy-2-methylphenylethynyl)phenyl]-1H-imidazol-2-ylaminehydrochloride

In a similar manner, 0.039 g (0.077 mmol) of2-tert-butoxycarbonylamino-5-[2-(4-methoxy-2-methylphenylethynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester afforded 0.024 g (92%) of5-[2-(4-methoxy-2-methylphenylethynyl)phenyl]-1H-imidazol-2-ylamine inits corresponding hydrochloride salt form as a tan amorphous solid: ¹HNMR (300 MHz, DMSO-d₆) δ 14.03 (s, 1H), 12.67 (s, 1H), 8.34 (d, 1H,J=8.1 Hz), 8.03 (d, 1H, J=8.1 Hz), 8.00 (br s, 2H), 7.66 (m, 3H), 7.51(m, 1H), 7.26 (d, 1H, J=8.1 Hz), 7.00 (d, 1H, J=2.7 Hz), 6.92 (dd, 1H,J=2.7, 8.1 Hz), 3.83 (s, 3H), 2.14 (s, 3H); ¹³C NMR (75 MHz, DMSO-d₆) δ159.3, 150.3, 137.5, 131.1, 129.8, 128.7, 128.1, 126.7, 125.5, 125.2,124.1, 124.0, 120.7, 119.1, 115.7, 111.6, 55.2, 19.9; HRMS (ESI) calcdfor C₁₉H₁₇N₃O (M⁺) 303.1372, found 303.1371.

5-[3-(3-dimethylaminoprop-1-ynyl)-phenyl]-1H-imidazol-2-ylaminedihydrochloride

In a similar manner, 0.036 g (0.082 mmol) of2-tert-butoxycarbonylamino-5-[3-(3-dimethylaminoprop-1-ynyl)phenyl]imidazole-1-carboxylicacid tert-butyl ester afforded 0.025 g (99%) of5-[3-(3-dimethylaminoprop-1-ynyl)-phenyl]-1H-imidazol-2-ylamine in itscorresponding dihydrochloride salt form as a yellow solid: ¹H NMR (300MHz, DMSO-d₆) δ 13.18 (s, 1H), 12.30 (s, 1H), 11.43 (s, 1H), 7.91 (s,1H), 7.67 (m, 1H), 7.50 (m, 5H), 4.33 (s, 2H), 2.86 (s, 6H); ¹³C NMR (75MHz, DMSO-d₆) δ 147.9, 131.0, 129.6, 128.5, 127.1, 125.4, 125.2, 121.6,110.6, 88.2, 79.7, 46.1, 41.6; HRMS (ESI) calcd for C₁₇H₁₃N₃ (M⁺)240.1375, found 240.1366.

1-(4-azidophenyl)-2-bromoethanone

4-azidobenzoic acid (2.71 g, 10.0 mmol) was dissolved in anhydrousdichloromethane (80 mL) and cooled to 0° C. Oxalyl chloride (4.40 mL,50.0 mmol) was added drop-wise followed by the addition of a catalyticamount of anhydrous DMF (0.01 mL). The reaction was allowed to warm toroom temperature over the course of 1 h and after that time the solutionwas evaporated to dryness. The crude acid chloride was dissolved inanhydrous dichloromethane (10 mL) added drop-wise to a 0° C. solution ofCH₂N₂ (50.0 mmol generated from Diazald®/KOH) in diethyl ether (150 mL).This solution was stirred at 0° C. for 1 h upon which the reaction wasquenched via the drop-wise addition of 48% solution of concentrated HBr(6.0 mL). The reaction mixture was diluted with dichloromethane (15 mL)and immediately washed with sat. NaHCO₃ (3×25 mL) and brine (2×25 mL)before being dried (MgSO₄), filtered and concentrated. The crude oil waspurified via flash column chromatography (10-50% EtOAc/Hexanes) toobtain the desired compound 1-(4-azidophenyl)-2-bromoethanone (3.84 g,96%) as a pale yellow solid: ¹H NMR (400 MHz, DMSO-d₆) δ 8.04 (m, 2H),7.28 (m, 2H), 4.90 (s, 2H); ¹³C NMR (100 MHz, DMSO-d₆) δ 190.4, 145.0,130.9, 130.5, 119.4, 33.8; HRMS (ESI) calcd for C₈H₆BrN₃O (M⁺) 238.9694,found 238.9688.

2-amino-5-(4-azidophenyl)imidazole-1-carboxylic acid tert-butyl ester

To a solution of 1-(4-azidophenyl)-2-bromoethanone (1.50 g, 6.24 mmol)in anhydrous DMF (20 mL) was added Boc-guanidine (3.00 g, 18.7 mmol).The reaction was stirred at ambient temperature for 48 hours upon whichthe mixture was partitioned between EtOAc (150 mL) and water (75 mL).The organic layer was successively washed with water (3×50 mL) and brine(2×50 mL) before being dried (Na₂SO₄) and evaporated to dryness. Theresulting crude oil was purified via flash column chromatography (10-50%EtOAc/Hexanes) to obtain the target2-amino-5-(4-azidophenyl)imidazole-1-carboxylic acid tert-butyl ester(1.84 g, 58%) as a yellow solid: ¹H NMR (300 MHz, DMSO-d₆) δ 7.77 (d,2H, J=8.1 Hz), 7.36 (s, 1H), 7.10 (dd, 2H, J=0.9, 8.1 Hz), 6.61 (br s,2H), 1.58 (s, 9H); ¹³C NMR (75 MHz, DMSO-d₆) δ 150.4, 148.9, 137.6,136.2, 130.5, 126.2, 119.1, 106.2, 84.7, 27.5; HRMS (ESI) calcd forC₈H₆BrN₃O (M⁺) 300.1335, found 300.1329.

5-(4-azido-phenyl)-2,2-tert-butoxycarbonylaminoimidazole-1-carboxylicacid tert-butyl ester

2-amino-5-(4-azidophenyl)imidazole-1-carboxylic acid tert-butyl ester(0.155 g, 0.516 mmol) and Boc anhydride (0.563 g, 2.58 mmol) weredissolved in anhydrous THF (8 mL) at room temperature. Triethylamine(1.05 mL, 7.74 mmol) and a catalytic amount of DMAP (0.003 g) were addedand the reaction was allowed to stir for 16 h. After that time, thesolution was poured into ethyl acetate (50 mL) and washed with 1N aq.HCl (3×10 mL), sat. aq. NaHCO₃ (3×10 mL), and brine (1×10 mL). Thecombined organics were dried (Na₂SO₄), filtered, and evaporated to yieldthe crude product which was purified by flash column chromatography(10-30% EtOAc/Hexanes) to give 0.200 g (78%) of the title compound5-(4-azido-phenyl)-2,2-tert-butoxycarbonylaminoimidazole-1-carboxylicacid tert-butyl ester as a tan foam: ¹H NMR (400 MHz, DMSO-d₆) δ 8.13(s, 1H), 7.89 (dd, 2H, J=0.8, 8.8 Hz), 7.14 (dd, 2H, J=0.8, 8.8 Hz),1.56 (s, 9H), 1.37 (s, 18H); ¹³C NMR (75 MHz, DMSO-d₆) δ 149.0, 145.9,138.5, 137.9, 137.3, 129.2, 126.4, 119.4, 114.0, 86.2, 83.4, 27.4, 27.3;HRMS (ESI) calcd for C₂₄H₃₂N₆O₆ (M⁺) 500.2383, found 500.2374.

2,2-tert-butoxycarbonylamino-5-[4-(3,5-difluorobenzoylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester

To a solution of anhydrous THF (5 mL) and 10% Pd/C (0.010 g) was charged5-(4-azido-phenyl)-2,2-tert-butoxycarbonylaminoimidazole-1-carboxylicacid tert-butyl ester (0.100 g, 0.200 mmol). Air was removed from thesystem and the reaction was back flushed with hydrogen. This process wasrepeated three times before setting the reaction under a hydrogenballoon at atmospheric pressure and temperature for 12 h. After thattime, the reaction was filtered to remove the catalyst. The filtrate wascooled to −78° C. and triethylamine (0.027 mL, 0.200 mmol) was addedprior to the drop-wise addition of 3,5-difluorobenzoyl chloride (0.035g, 0.200 mmol) diluted in anhydrous dichloromethane (0.50 mL). Thereaction was stirred at −78° C. for 30 mins and then quenched withmethanol (1 mL) before being concentrated under reduced pressure andpurified by flash column chromatography (10-40% EtOAc/Hexanes) to obtainthe title compound2,2-tert-butoxycarbonylamino-5-[4-(3,5-difluorobenzoylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester (0.094 g, 77%) as a pale yellow solid: ¹H NMR (300MHz, DMSO-d₆) δ 10.42 (s, 1H), 8.08 (s, 1H), 7.82 (m, 4H), 7.69 (m, 2H),7.54 (m, 1H), 1.57 (s, 9H), 1.38 (s, 18H); ¹³C NMR (100 MHz, CD₃OD) δ165.8, 163.2, 151.0, 147.7, 140.1, 139.6, 130.0, 126.7, 122.4, 114.7,111.9, 108.1, 88.2, 85.7, 28.3; HRMS (ESI) calcd for C₃₁H₃₆F₂N₄O₇ (M⁺)614.2552, found 614.2554.

2,2-tert-butoxycarbonylamino-5-[4-(4-chlorobutyrylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.200 mmol) of5-(4-azido-phenyl)-2,2-tert-butoxycarbonylaminoimidazole-1-carboxylicacid tert-butyl ester afforded 0.074 g (64%) of2,2-tert-butoxycarbonylamino-5-[4-(4-chlorobutyrylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a tan solid: ¹H NMR (300 MHz, DMSO-d₆) δ 10.04(s, 1H), 8.00 (s, 1H), 7.77 (d, 2H, J=8.4 Hz), 7.60 (d, 2H, J=8.4 Hz),3.69 (t, 2H, J=6.3 Hz), 2.48 (m, 2H), 2.00 (m, 2H), 1.55 (s, 9H), 1.36(s, 18H); ¹³C NMR (100 MHz, CD₃OD) δ 173.3, 151.0, 147.7, 140.0, 129.2,126.7, 121.4, 114.4, 88.2, 85.6, 88.2, 85.6, 45.3, 35.0, 29.7, 28.3;HRMS (ESI) calcd for C₂₈H₃₉ClN₄O₇ (M⁺) 578.2507, found 578.2510.

4-(2-amino-3H-imidazol-4-yl)phenyl]-3,5-difluorobenzamide hydrochloride

2,2-tert-butoxycarbonylamino-5-[4-(3,5-difluorobenzoylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester (0.046 g, 0.078 mmol) was dissolved in anhydrousdichloromethane (5 mL) and cooled to 0° C. Trifluoroacetic acid (0.50mL) was added drop-wise while the reaction continued to stir at 0° C.Upon completion, the reaction was allowed to warm to room temperatureover the course of 12 h. Toluene (2 mL) was added and the reaction wasevaporated to dryness. The crude TFA salt was then dissolved indichloromethane (3 mL) and a 2M solution of HCl in diethyl ether (0.10mL) was added. The solution was again concentrated under reducedpressure and the resulting product triturated with cold diethyl ether (5mL) to afford 0.026 g (99%) of4-(2-amino-3H-imidazol-4-yl)phenyl]-3,5-difluorobenzamide in itscorresponding hydrochloride salt form as a white solid: ¹H NMR (300 MHz,DMSO-d₆) δ 12.94 (s, 1H), 12.14 (s, 1H), 10.57 (s, 1H), 7.85 (d, 2H,J=8.4 Hz), 7.72 (m, 2H), 7.66 (d, 2H, J=8.4 Hz), 7.48 (br s, 2H), 7.35(s, 1H); ¹³C NMR (100 MHz, CD₃OD) δ 165.8, 149.6, 140.0, 139.8, 128.8,126.2, 125.3, 122.6, 112.2, 111.9, 109.7, 108.1; HRMS (ESI) calcd forC₁₆H₁₂F₂N₄O (M⁺) 314.0979, found 314.0977.

4-(2-amino-3H-imidazol-4-yl)phenyl]-4-chlorobutyramide hydrochloride

In a similar manner, 0.051 g (0.089 mmol) of2,2-tert-butoxycarbonylamino-5-[4-(4-chlorobutyrylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester afforded 0.026 g (96%) of4-(2-amino-3H-imidazol-4-yl)phenyl]-4-chlorobutyramide in itscorresponding hydrochloride salt form as a tan solid: ¹H NMR (300 MHz,DMSO-d₆) δ 12.82 (s, 1H), 12.07 (s, 1H), 10.19 (s, 1H), 7.65 (d, 2H,J=9.0 Hz), 7.57 (d, 2H, J=8.7 Hz), 7.45 (br s, 2H), 7.29 (s, 1H), 3.70(d, 2H, J=6.3 Hz), 2.47 (m, 2H), 2.03 (m, 2H); ¹³C NMR (100 MHz, CD₃OD)δ 173.5, 148.5, 140.4, 129.0, 126.3, 124.6, 121.6, 109.4, 45.3, 35.0,29.7; HRMS (ESI) calcd for C₁₃H₁₅ClN₄O (M⁺) 278.0934, found 278.0931.

Cyclopropanecarboxylic acid [4-(2-amino-3H-imidazol-4-yl)phenyl]amidehydrochloride

In a similar manner, 0.060 g (0.111 mmol) of2,2-tert-butoxycarbonylamino-5-[4-(cyclopropanecarbonylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester afforded 0.030 g (99%) of cyclopropanecarboxylicacid [4-(2-amino-3H-imidazol-4-yl)phenyl]amide in its correspondinghydrochloride salt form as a tan amorphous solid: ¹H NMR (400 MHz,DMSO-d₆) δ 12.78 (s, 1H), 12.05 (s, 1H), 10.41 (s, 1H), 7.67 (d, 2H,J=8.0 Hz), 7.56 (d, 2H, J=8.0 Hz), 7.45 (br s, 2H), 7.28 (s, 1H), 1.84(m, 1H), 0.80 (m, 4H); ¹³C NMR (100 MHz, CD₃OD) δ 175.2, 163.2, 149.3,140.7, 126.3, 124.4, 121.4, 109.4, 15.8, 8.29; HRMS (ESI) calcd forC₁₃H₁₄N₄O (M⁺) 242.1168, found 242.1160.

4-(2-amino-3H-imidazol-4-yl)phenyl-2-(4-methoxyphenyl)acetamidehydrochloride

In a similar manner, 0.041 g (0.066 mmol) of2,2-tert-butoxycarbonylamino-5-{4-[2-(4-methoxyphenyl)acetylamino]phenyl}imidazole-1-carboxylicacid tert-butyl ester afforded 0.023 g (99%) of4-(2-amino-3H-imidazol-4-yl)phenyl-2-(4-methoxyphenyl)acetamide in itscorresponding hydrochloride salt form as a tan solid: ¹H NMR (300 MHz,DMSO-d₆) δ 12.80 (s, 1H), 12.06 (s, 1H), 10.36 (s, 1H), 7.65 (d, 2H,J=8.7 Hz), 7.56 (d, 2H, J=8.7 Hz), 7.44 (br s, 2H), 7.24 (m, 3H), 6.86(d, 2H, J=8.7 Hz), 3.71 (s, 3H), 3.57 (s, 2H); ¹³C NMR (100 MHz, CD₃OD)δ 160.3, 149.3, 140.3, 131.2, 129.2, 128.8, 127.3, 126.2, 124.7, 121.6,115.1, 109.7, 55.8, 43.9; HRMS (ESI) calcd for C₁₈H₁₈N₄O₂ (M⁺) 322.1430,found 322.1428.

4-(2-amino-3H-imidazol-4-yl)phenyl]-4-pentylbenzamide hydrochloride

In a similar manner, 0.085 g (0.131 mmol) of2,2-tert-butoxycarbonylamino-5-[4-(4-pentylbenzoylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester afforded 0.050 g (99%) of4-(2-amino-3H-imidazol-4-yl)phenyl]-4-pentylbenzamide in itscorresponding hydrochloride salt form as a tan amorphous solid: ¹H NMR(400 MHz, DMSO-d₆) δ 13.09 (s, 1H), 12.23 (s, 1H), 10.36 (s, 1H), 7.86(m, 4H), 7.64 (d, 2H, J=8.8 Hz), 7.52 (br s, 2H), 7.33 (m, 3H), 2.62 (t,2H, J=7.6 Hz), 1.60 (m, 2H), 1.28 (m, 4H), 0.83 (t, 3H, J=5.7 Hz); ¹³CNMR (100 MHz, CD₃OD) δ 167.7, 147.6, 139.2, 133.6, 131.0, 129.8, 129.6,128.9, 126.2, 124.9, 122.6, 108.2, 35.6, 31.4, 30.9, 22.4, 13.2; HRMS(ESI) calcd for C₂₁H₂₄N₄O (M⁺) 348.1950, found 348.1939.

2-amino-5-(4-heptanoylaminophenyl)imidazole-1-carboxylic acid tert-butylester

To a solution of anhydrous THF (5 mL) and 10% Pd/C (0.010 g) was charged2-amino-5-(4-azidophenyl)imidazole-1-carboxylic acid tert-butyl ester(0.100 g, 0.332 mmol). Air was removed from the system and the reactionwas back flushed with hydrogen. This process was repeated three timesbefore setting the reaction under a hydrogen balloon at atmosphericpressure and temperature for 12 h. After that time, the reaction wasfiltered to remove the catalyst. The filtrate was cooled to −78° C. andtriethylamine (0.046 mL, 0.332 mmol) was added prior to the drop-wiseaddition of heptanoyl chloride (0.049 g, 0.332 mmol) diluted inanhydrous dichloromethane (0.50 mL). The reaction was stirred at −78° C.for 30 mins and then quenched with methanol (1 mL) before beingconcentrated under reduced pressure and purified by flash columnchromatography (10-60% EtOAc/Hexanes) to obtain the title compound2-amino-5-(4-heptanoylaminophenyl)imidazole-1-carboxylic acid tert-butylester (0.124 g, 97%) as a yellow solid: ¹H NMR (300 MHz, DMSO-d₆) δ10.08 (s, 1H), 8.14 (br s, 2H), 7.67 (m, 4H), 7.56 (s, 1H), 2.32 (t, 2H,J=7.5 Hz), 1.49 (m, 11H), 1.28 (br s, 6H), 0.87 (t, 2H, J=6.0 Hz); ¹³CNMR (75 MHz, DMSO-d₆) δ 171.4, 147.5, 139.1, 126.4, 124.7, 122.2, 119.2,119.0, 111.4, 108.4, 36.4, 31.0, 28.3, 27.4, 25.0, 21.9, 13.9; HRMS(ESI) calcd for C₂₁H₃₀N₄O₃ (M⁺) 386.2318, found 386.2312.

2-amino-5-[4-(6-bromohexanoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester

In a similar manner, 0.100 g (0.332 mmol) of2-amino-5-(4-azidophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.124 g (83%) of2-amino-5-[4-(6-bromohexanoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester as a pale yellow foam: ¹H NMR (300 MHz, DMSO-d₆) δ 9.89(s, 1H), 7.63 (d, 2H, J=8.4 Hz), 7.57 (d, 2H, J=8.0 Hz), 7.24 (s, 1H),6.58 (br s, 2H), 3.53 (m, 2H), 2.30 (t, 2H, J=7.2 Hz), 1.82 (tt, 2H,J=6.8, 13.6 Hz), 1.57 (m, 11H), 1.41 (m, 2H); ¹³C NMR (100 MHz, DMSO-d₆)δ 171.6, 151.1, 149.9, 137.7, 129.6, 126.0, 120.3, 106.3, 85.7, 37.8,34.2, 32.9, 28.5, 28.2, 25.1; HRMS (ESI) calcd for C₂₀H₂₇N₄O₃ (M⁺)450.1267, found 450.1258.

2-amino-5-[4-(cyclobutanecarbonylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.100 g (0.332 mmol) of2-amino-5-(4-azidophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.088 g (75%) of2-amino-5-[4-(cyclobutanecarbonylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a white foam: ¹H NMR (400 MHz, DMSO-d₆) δ 9.73(s, 1H), 7.63 (d, 2H, J=8.0 Hz), 7.57 (d, 2H, J=8.0 Hz), 7.25 (s, 1H),6.58 (br s, 2H), 3.21 (m, 1H), 2.21 (m, 2H), 2.10 (m, 2H), 1.94 (m, 1H),1.81 (m, 1H), 1.58 (s, 9H); ¹³C NMR (100 MHz, DMSO-d₆) δ 173.4, 150.7,149.6, 137.5, 129.2, 125.9, 125.7, 119.8, 105.9, 85.4, 41.0, 28.2, 25.5,18.2; HRMS (ESI) calcd for C₁₉H₂₄N₄O₃ (M⁺) 356.1848, found 356.1844.

2-amino-5-[4-(3-phenylacryloylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester

In a similar manner, 0.100 g (0.332 mmol) of2-amino-5-(4-azidophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.078 g (58%) of2-amino-5-[4-(3-phenylacryloylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester as a tan amorphous solid: ¹H NMR (400 MHz, DMSO-d₆) δ10.25 (s, 1H), 7.68 (m, 7H), 7.43 (m, 3H), 7.28 (s, 1H), 6.83 (d, 1H,J=15.2 Hz), 6.60 (br s, 2H), 1.58 (s, 9H); ¹³C NMR (100 MHz, CDCl₃) δ164.3, 150.7, 149.6, 142.4, 137.6, 137.4, 134.9, 130.1, 129.5, 129.0,128.1, 125.9, 121.2, 120.2, 106.1, 85.4, 28.2; HRMS (ESI) calcd forC₂₃H₂₄N₄O₃ (M⁺) 404.1848, found 484.1850.

Heptanoic acid [4-(2-amino-3H-imidazol-4-yl)phenyl]amide hydrochloride

2-amino-5-(4-heptanoylaminophenyl)imidazole-1-carboxylic acid tert-butylester (0.089 g, 0.230 mmol) was dissolved in anhydrous dichloromethane(5 mL) and cooled to 0° C. Trifluoroacetic acid (0.50 mL) was addeddrop-wise while the reaction continued to stir at 0° C. Upon completion,the reaction was allowed to warm to room temperature over the course of12 h. Toluene (2 mL) was added and the reaction was evaporated todryness. The crude TFA salt was then dissolved in dichloromethane (3 mL)and a 2M solution of HCl in diethyl ether (0.10 mL) was added. Thesolution was again concentrated under reduced pressure and the resultingproduct triturated with cold diethyl ether (5 mL) to afford 0.072 g(97%) of the desired compound heptanoic acid[4-(2-amino-3H-imidazol-4-yl)phenyl]amide in its correspondinghydrochloride form as a yellow amorphous solid: ¹H NMR (300 MHz,DMSO-d₆) δ 12.80 (s, 1H), 12.05 (s, 1H), 10.08 (s, 1H), 7.67 (d, 2H,J=8.1 Hz), 7.59 (d, 2H, J=8.1 Hz), 7.43 (br s, 2H), 7.27 (s, 1H), 2.31(t, 2H, J=6.8 Hz), 1.58 (m, 2H), 1.27 (m, 6H), 0.86 (t, 3H, J=5.7 Hz);¹³C NMR (75 MHz, DMSO-d₆) δ 172.1, 148.2, 139.8, 127.1, 125.4, 123.0,119.9, 109.1, 37.1, 31.7, 29.0, 25.7, 22.7, 14.6; HRMS (ESI) calcd forC₁₆H₂₂N₄O (M⁺) 286.1794, found 286.1788.

Cyclobutanecarboxylic acid [4-(2-amino-3H-imidazol-4-yl)phenyl]amidehydrochloride

In a similar manner, 0.060 g (0.168 mmol) of2-amino-5-[4-(cyclobutanecarbonylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester afforded 0.048 g (98%) of cyclobutanecarboxylicacid [4-(2-amino-3H-imidazol-4-yl)phenyl]amide in its correspondinghydrochloride salt form as a yellow solid: ¹H NMR (400 MHz, DMSO-d₆) δ12.76 (s, 1H), 12.03 (s, 1H), 9.92 (s, 1H), 7.67 (d, 2H, J=9.2 Hz), 7.56(d, 2H, J=8.4 Hz), 7.42 (br s, 2H), 7.28 (s, 1H), 3.22 (m, 1H), 2.19 (m,2H), 2.11 (m, 2H), 1.92 (m, 1H), 1.81 (m, 1H); ¹³C NMR (100 MHz, CD₃OD)δ 176.4, 149.3, 140.6, 128.9, 126.2, 124.4, 121.6, 109.4, 41.8, 26.2,19.1; HRMS (ESI) calcd for C₁₄H₁₆N₄O (M⁺) 256.1324, found 256.1321.

6-bromohexanoic acid [4-(2-amino-3H-imidazol-4-yl)phenyl]amidehydrochloride

In a similar manner, 0.046 g (0.101 mmol) of2-amino-5-[4-(6-bromohexanoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester afforded 0.039 g (99%) of 6-bromohexanoic acid[4-(2-amino-3H-imidazol-4-yl)phenyl]amide in its correspondinghydrochloride salt form as a yellow solid: ¹H NMR (400 MHz, DMSO-d₆) δ12.80 (s, 1H), 12.05 (s, 1H), 10.10 (s, 1H), 7.65 (d, 2H, J=8.7 Hz),7.56 (d, 2H, J=8.7 Hz), 7.44 (br s, 2H), 7.28 (s, 1H), 3.54 (t, 2H,J=6.4 Hz), 2.33 (t, 2H, J=7.2 Hz), 1.82 (tt, 2H, J=6.8, 14.0 Hz), 1.61(tt, 2H, J=6.8, 14.8 Hz), 1.41 (tt, 2H, J=7.6, 14.8 Hz); ¹³C NMR (100MHz, CD₃OD) δ 175.0, 148.1, 140.8, 129.3, 126.6, 124.9, 121.9, 109.8,38.2, 34.6, 34.1, 29.3, 26.4; HRMS (ESI) calcd for C₁₅H₁₉BrN₄O (M⁺)350.0742, found 350.0739.

4-(2-amino-3H-imidazol-4-yl)phenyl-3-phenylacrylamide hydrochloride

In a similar manner, 0.061 g (0.151 mmol) of2-amino-5-[4-(3-phenylacryloylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester afforded 0.050 g (98%) of4-(2-amino-3H-imidazol-4-yl)phenyl-3-phenylacrylamide in itscorresponding hydrochloride salt form as a yellow solid: ¹H NMR (300MHz, DMSO-d₆) δ 12.80 (s, 1H), 12.06 (s, 1H), 10.49 (s, 1H), 7.77 (d,2H, J=8.7 Hz), 7.62 (m, 5H), 7.43 (m, 5H), 7.32 (s, 1H), 6.87 (d, 1H,J=15.9 Hz); ¹³C NMR (100 MHz, CD₃OD) δ 166.8, 149.3, 143.2, 140.5,136.3, 131.3, 130.2, 129.1, 128.9, 126.3, 124.7, 122.2, 121.6, 109.5;HRMS (ESI) calcd for C₁₈H₁₆N₄O (M⁺) 304.1324, found 304.1319.

3-(2-bromoacetyl)phenyl carbamic acid benzyl ester

3-acetylphenyl carbamic acid benzyl ester (3.50 g, 13.0 mmol) wasdissolved in diethyl ether (70 mL) and THF (20 mL) and brought to 0° C.

A catalytic amount of aluminum (III) chloride (0.010 g) was addedfollowed by the drop-wise addition of bromine (0.731 mL, 14.3 mmol) indiethyl ether (10 mL). The reaction was kept at 0° C. for 30 mins beforebeing allowed to warm to room temperature on its own accord. Sat. NaHCO₃(30 mL) was added to quench the reaction and the solution was pouredinto ethyl acetate (30 mL). The combined organics were washed with sat.NaHCO₃ (3×30 mL) and brine (1×30 mL) before being dried (Na₂SO₄),filtered, and evaporated to dryness. Trituration of the resulting crudeproduct with a 2:1 mixture of dichloromethane/diethyl ether (6 mL)followed by filtration afforded 2.57 g (57%) of 3-(2-bromoacetyl)phenylcarbamic acid benzyl ester as a light pink solid: ¹H NMR (400 MHz,DMSO-d₆) δ 10.04 (s, 1H), 8.11 (s, 1H), 7.69 (m, 2H), 7.43 (m, 6H), 5.18(s, 2H), 4.88 (s, 2H); ¹³C NMR (100 MHz, DMSO-d₆) δ 201.5, 163.4, 149.8,146.5, 144.6, 139.4, 138.5, 138.2, 133.3, 131.9, 127.6, 126.7, 76.0,44.1; HRMS (ESI) calcd for C₁₆H₁₄BrNO₃ (M⁺) 347.0157, found 347.0151.

2-amino-5-(3-benzyloxycarbonyl aminophenyl)imidazole-1-carboxylic acidtert-butyl ester

3-(2-bromoacetyl)phenyl carbamic acid benzyl ester (2.94 g, 8.43 mmol)and Boc-guanidine (4.02 g, 25.3 mmol) were dissolved in DMF (25 mL) andallowed to stir at room temperature. After 24 h the DMF was removedunder reduced pressure and the residue taken up in ethyl acetate (50 mL)and washed with water (3×25 mL) and brine (25 mL) before being dried(Na₂SO₄), filtered, and evaporated to dryness. The resulting crude oilwas purified by flash column chromatography (10-40% EtOAc/Hexanes) toobtain the pure product as an oil. This oil was triturated with coldhexanes (15 mL) and filtered to afford 1.40 g (48%) of2-amino-5-(3-benzyloxycarbonyl aminophenyl)imidazole-1-carboxylic acidtert-butyl ester as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ 9.75 (s,1H), 7.89 (s, 1H), 7.22-7.42 (m, 9H), 6.62 (s, 2H), 5.15 (s, 2H), 1.58(s, 9H); ¹³C NMR (100 MHz, DMSO-d₆) δ 153.4, 150.4, 148.9, 139.2, 137.0,136.7, 134.0, 128.7, 128.4, 128.0, 119.0, 117.0, 114.7, 106.0, 84.7,65.7, 27.5; HRMS (ESI) calcd for C₂₂H₂₄N₄O₄ (M⁺) 408.1798, found408.1796.

2-amino-5-(3-aminophenyl)imidazole-1-carboxylic acid tert-butyl ester

To a solution of absolute ethanol (50 mL) and 10% Pd/C (0.150 g) wascharged 2-amino-5-(3-benzyloxycarbonylaminophenyl)imidazole-1-carboxylic acid tert-butyl ester (1.40 g, 3.42mmol). Air was removed from the system and the reaction was back flushedwith hydrogen. This process was repeated three times before setting thereaction under 35 psi of hydrogen using a Parr-Shaker apparatus at roomtemperature for 16 h. After that time, the reaction was filtered toremove the catalyst and the filtrate evaporated to deliver 0.928 g (99%)of 2-amino-5-(3-aminophenyl)imidazole-1-carboxylic acid tert-butyl esteras a white solid. ¹H NMR (300 MHz, DMSO-d₆) δ 7.11 (s, 1H), 6.95 (m,2H), 6.85 (dd, 1H, J=0.9, 7.5 Hz), 6.53 (br s, 2H), 6.42 (m, 1H), 5.01(br s, 2H), 1.57 (s, 9H); ¹³C NMR (100 MHz, DMSO-d₆) δ 150.2, 150.0,148.6, 137.8, 133.8, 128.8, 112.9, 112.7, 110.4, 105.3, 84.5, 27.6; HRMS(ESI) calcd for C₁₄H₁₈N₄O₂ (M⁺) 274.1430, found 274.1425.

5-[3-(adamantane-1-carbonylamino)phenyl]-2-aminoimidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(3-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.102 g (86%) of5-[3-(adamantane-1-carbonylamino)phenyl]-2-aminoimidazole-1-carboxylicacid tert-butyl ester as a white solid: ¹H NMR (400 MHz, DMSO-d₆) δ 9.15(s, 1H), 8.00 (s, 1H), 7.56 (d, 1H, J=7.6 Hz), 7.38 (d, 1H, J=7.6 Hz),7.22 (m, 2H), 6.63 (br s, 2H), 2.01 (br s, 4H), 1.92 (br s, 6H), 1.70(br s, 6H), 1.58 (s, 9H); ¹³C NMR (100 MHz, DMSO-d₆) δ 175.9, 150.4,148.9, 139.5, 137.1, 133.5, 128.4, 119.6, 118.9, 116.7, 105.8, 84.7,40.9, 38.3, 36.0, 27.7, 27.5; HRMS (ESI) calcd for C₂₅H₃₂N₄O₃ (M⁺)436.2474, found 436.2468.

2-amino-5-{3-[(benzo[1,3]dioxole-5-carbonyl)-amino]phenyl}imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(3-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.091 g (79%) of2-amino-5-{3-[(benzo[1,3]dioxole-5-carbonyl)-amino]phenyl}imidazole-1-carboxylicacid tert-butyl ester as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ10.07 (s, 1H), 8.12 (m, 1H), 7.54-7.65 (m, 3H), 7.44 (m, 1H), 7.29 (t,1H, J=7.8 Hz), 7.26 (s, 1H), 7.04 (d, 1H, J=8.4 Hz), 6.63 (br s, 2H),6.14 (s, 2H), 1.59 (s, 9H); ¹³C NMR (100 MHz, DMSO-d₆) δ 164.3, 150.4,150.0, 148.9, 147.4, 139.4, 139.3, 137.0, 133.7, 128.7, 128.6, 122.9,120.0, 118.9, 116.8, 107.9, 107.7, 106.0, 101.8, 84.7, 27.5; HRMS (ESI)calcd for C₂₂H₂₂N₄O₅ (M⁺) 422.1590, found 422.1585.

2-amino-5-[3-(4-pentylbenzoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(3-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.094 g (77%) of2-amino-5-[3-(4-pentylbenzoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ 10.18 (s,1H), 8.12 (m, 1H), 7.90 (d, 2H, J=8.4 Hz), 7.64 (d, 1H, J=7.8 Hz), 7.45(d, 1H, J=7.8 Hz), 7.31 (m, 4H), 6.64 (br s, 2H), 2.65 (t, 2H, J=7.8Hz), 1.66 (m, 2H), 1.59 (s, 9H), 1.29 (m, 4H), 0.87 (t, 3H, J=5.1 Hz);¹³C NMR (100 MHz, CDCl₃) δ 165.9, 150.7, 149.6, 147.5, 138.6, 137.3,134.2, 132.5, 129.5, 127.3, 121.1, 119.1, 116.7, 106.8, 85.5, 36.0,31.6, 31.1, 28.2, 22.7, 14.2; HRMS (ESI) calcd for C₂₆H₃₂N₄O₃ (M⁺)448.2474, found 448.2472.

2-amino-5-{3-[(thiophene-2-carbonyl)amino]phenyl}imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(3-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.061 g (59%) of2-amino-5-{3-[(thiophene-2-carbonyl)amino]phenyl}imidazole-1-carboxylicacid tert-butyl ester as a white foam: ¹H NMR (400 MHz, CDCl₃) δ 7.96(s, 1H), 7.86 (t, 1H, J=1.6 Hz), 7.70 (m, 1H), 7.61 (dd, 1H, J=1.2, 4.0Hz), 7.53 (dd, 1H, J=1.2, 4.8 Hz), 7.45 (m, 1H), 7.34 (t, 1H, J=7.6 Hz),7.14 (s, 1H), 7.10 (m, 1H), 5.93 (br s, 2H), 1.63 (s, 9H); ¹³C NMR (100MHz, CDCl₃) δ 160.1, 150.6, 149.6, 139.6, 138.2, 137.2, 134.3, 130.9,129.5, 128.6, 128.0, 121.3, 119.2, 116.7, 106.9, 85.5, 28.2; HRMS (ESI)calcd for C₁₉H₂₀N₄O₃S (M⁺) 384.1256, found 384.1250.

2-amino-5-[3-(3-cyclopentylpropionylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(3-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.085 g (79%) of2-amino-5-[3-(3-cyclopentylpropionylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a white foam: ¹H NMR (300 MHz, DMSO-d₆) δ 9.91(s, 1H), 7.96 (s, 1H), 7.45 (d, 1H, J=8.1 Hz), 7.37 (d, 1H, J=8.1 Hz),7.27 (m, 2H), 6.98 (br s, 2H), 2.31 (t, 2H, J=7.5 Hz), 1.75 (m, 2H),1.51 (m, 16H), 1.10 (m, 2H); ¹³C NMR (100 MHz, CDCl₃) δ 172.0, 150.5,149.1, 138.8, 135.0, 132.1, 129.6, 120.8, 119.4, 116.2, 106.6, 86.6,39.9, 37.2, 32.7, 32.0, 28.2, 25.3; HRMS (ESI) calcd for C₂₂H₃₀N₄O₃ (M⁺)398.2318, found 398.2311.

2-amino-5-{3-[(furan-2-carbonyl)amino]phenyl}imidazole-1-carboxylic acidtert-butyl ester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(3-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.058 g (58%) of2-amino-5-{3-[(furan-2-carbonyl)amino]phenyl}imidazole-1-carboxylic acidtert-butyl ester as a white foam: ¹H NMR (300 MHz, DMSO-d₆) δ 10.17 (s,1H), 8.11 (m, 1H), 7.94 (m, 1H), 7.62 (m, 1H), 7.45 (m, 1H), 7.35 (dd,1H, J=1.2, 3.6 Hz), 7.30 (t, 1H, J=8.1 Hz), 7.27 (s, 1H), 6.71 (m, 1H),6.63 (br s, 2H), 1.51 (s, 9H); ¹³C NMR (100 MHz, CDCl₃) δ 156.3, 150.8,149.6, 148.0, 144.4, 137.8, 137.3, 134.3, 129.5, 121.2, 118.2, 116.4,115.3, 112.7, 106.8, 85.4, 28.2; HRMS (ESI) calcd for C₁₉H₂₀N₄O₄ (M⁺)368.1485, found 368.1479.

2-amino-5-[3-(2-ethylhexanoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(3-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.088 g (81%) of2-amino-5-[3-(2-ethylhexanoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ 9.87 (s,1H), 8.00 (s, 1H), 7.47 (m, 1H), 7.38 (m, 1H), 7.24 (m, 2H), 6.70 (br s,2H), 2.28 (tt, 1H, J=4.5, 9.0 Hz), 1.23-1.58 (m, 17H), 0.85 (m, 6H); ¹³CNMR (100 MHz, CDCl₃) δ 174.7, 150.5, 149.7, 138.5, 136.8, 133.7, 129.5,120.9, 119.0, 116.2, 106.8, 85.8, 51.2, 32.9, 30.1, 28.2, 26.4, 23.0,14.2, 12.3; HRMS (ESI) calcd for C₂₂H₃₂N₄O₃ (M⁺) 400.2474, found400.2472.

2-amino-5-[3-(6-bromohexanoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(3-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.090 g (73%) of2-amino-5-[3-(6-bromohexanoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ 9.89 (s,1H), 7.96 (s, 1H), 7.35 (d, 1H, J=8.1 Hz), 7.26 (d, 1H, J=7.8 Hz), 7.22(s, 1H), 6.61 (br s, 2H), 3.54 (t, 2H, J=5.4 Hz), 2.31 (m, 2H), 1.83 (m,2H), 1.55 (s, 9H), 1.42 (m, 2H); ¹³C NMR (100 MHz, CD₃OD) δ 171.2,150.6, 149.6, 138.4, 137.4, 134.2, 129.5, 121.1, 118.9, 116.3, 107.0,85.6, 37.6, 33.8, 32.7, 28.2, 28.0, 24.8; HRMS (ESI) calcd for C₁₄H₁₇N₄O(M⁺) 256.1324, found 256.1319.

2-amino-5-(3-heptanoylaminophenyl)imidazole-1-carboxylic acid tert-butylester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(3-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.075 g (69%) of2-amino-5-(3-heptanoylaminophenyl)imidazole-1-carboxylic acid tert-butylester as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ 9.87 (s, 1H), 7.96(s, 1H), 7.37 (d, 1H, J=7.5 Hz), 7.35 (d, 1H, J=6.9 Hz), 7.23 (s, 1H),6.61 (br s, 2H), 2.27 (t, 2H, J=7.5 Hz), 1.59 (s, 9H), 1.27 (m, 8H),0.85 (m, 3H); ¹³C NMR (100 MHz, CD₃OD) δ 171.7, 150.7, 149.6, 138.5,137.3, 134.1, 129.4, 121.0, 118.9, 116.3, 106.8, 85.6, 38.1, 31.8, 29.1,28.2, 25.8, 22.7, 14.2; HRMS (ESI) calcd for C₂₁H₃₀N₄O₃ (M⁺) 386.2318,found 386.2313.

2-amino-5-[3-(4-hexylbenzoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(3-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.086 g (68%) of2-amino-5-[3-(4-hexylbenzoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester as a white foam: ¹H NMR (300 MHz, DMSO-d₆) δ 10.17 (s,1H), 8.15 (s, 1H), 7.89 (d, 2H, J=8.1), 7.65 (m, 1H), 7.45 (m, 1H), 7.34(d, 2H, J=8.1 Hz), 7.30 (s, 1H), 7.26 (s, 1H), 6.64 (br s, 2H), 2.65 (m,2H), 1.59 (s, 9H), 1.27 (m, 8H), 0.85 (m, 3H); ¹³C NMR (100 MHz, CD₃OD)δ 168.9, 152.8, 150.7, 148.6, 140.3, 138.1, 134.8, 133.7, 130.9, 130.1,129.5, 122.3, 121.4, 118.8, 107.9, 86.9, 37.0, 33.0, 32.5, 30.2, 28.3,23.8, 14.6; HRMS (ESI) calcd for C₂₇H₃₄N₄O₃ (M⁺) 462.2631, found462.2624.

2-amino-5-[3-(cyclohexanecarbonylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(3-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.088 g (82%) of2-amino-5-[3-(cyclohexanecarbonylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a white foam: ¹H NMR (300 MHz, DMSO-d₆) δ 9.72(s, 1H), 7.97 (s, 1H), 7.50 (d, 1H, J=8.1 Hz), 7.40 (d, 1H, J=5.6 Hz),7.26 (m, 1H), 6.61 (br s, 2H), 3.30 (m, 1H), 2.19 (m, 4H), 2.07 (m, 4H),1.90 (m, 2H), 1.55 (s, 9H); ¹³C NMR (100 MHz, CD₃OD) δ 174.6, 150.9,149.5, 138.6, 137.1, 133.9, 129.4, 120.9, 118.9, 116.3, 106.7, 85.6,43.8, 29.9, 28.2, 26.1, 25.9; HRMS (ESI) calcd for C₂₁H₂₈N₄O₃ (M⁺)384.2161, found 384.2158.

2-amino-5-[3-(2-nitrobenzoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(3-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.095 g (83%) of2-amino-5-[3-(2-nitrobenzoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester as a white foam: ¹H NMR (300 MHz, DMSO-d₆) δ 10.67 (s,1H), 8.12 (m, 1H), 8.11 (s, 1H), 7.85 (m, 1H), 7.80 (m, 2H), 7.48 (m,2H), 7.35 (m, 1H), 7.28 (s, 1H), 6.64 (br s, 2H), 1.55 (s, 9H); ¹³C NMR(100 MHz, CD₃OD) δ 167.5, 152.9, 150.7, 147.8, 140.1, 138.2, 135.3,135.2, 134.4, 132.1, 130.3, 125.7, 122.7, 120.7, 118.2, 108.0, 86.9;HRMS (ESI) calcd for C₂₁H₂₁N₅O₅ (M⁺) 423.1543, found 423.1564.

3-(2-amino-3H-imidazol-4-yl)phenyl]-2-nitrobenzamide hydrochloride

2-amino-5-[3-(2-nitrobenzoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester (0.070 g, 0.165 mmol) was dissolved in anhydrousdichloromethane (5 mL) and cooled to 0° C. Trifluoroacetic acid (0.50mL) was added drop-wise while the reaction continued to stir at 0° C.Upon completion, the reaction was allowed to warm to room temperatureover the course of 12 h. Toluene (2 mL) was added and the reaction wasevaporated to dryness. The crude TFA salt was then dissolved indichloromethane (3 mL) and a 2M solution of HCl in diethyl ether (0.10mL) was added. The solution was again concentrated under reducedpressure and the resulting product triturated with cold diethyl ether (5mL) to afford 0.058 g (98%) of the desired compound3-(2-amino-3H-imidazol-4-yl)phenyl]-2-nitrobenzamide in itscorresponding hydrochloride salt form as a yellow foam: ¹H NMR (300 MHz,DMSO-d₆) δ 12.82 (s, 1H), 12.17 (s, 1H), 10.85 (s, 1H), 8.17 (m, 1H),8.00 (s, 1H), 7.83 (m, 1H), 7.79 (m, 2H), 7.42-7.52 (m, 5H), 7.32 (s,1H); ¹³C NMR (100 MHz, CD₃OD) δ 165.8, 148.0, 147.0, 139.3, 134.1,132.8, 131.0, 129.8, 129.0, 128.5, 127.0, 124.5, 120.8, 120.4, 116.3,109.0; HRMS (ESI) calcd for C₁₆H₁₃N₅O₃ (M⁺) 323.1018, found 323.1022.

6-bromohexanoic acid [3-(2-amino-3H-imidazol-4-yl)phenyl]amidehydrochloride

In a similar manner, 0.050 g (0.110 mmol) of2-amino-5-[3-(6-bromohexanoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester afforded 0.041 g (98%) of 6-bromohexanoic acid[3-(2-amino-3H-imidazol-4-yl)phenyl]amide in its correspondinghydrochloride salt form as a yellow solid: ¹H NMR (300 MHz, DMSO-d₆) δ12.79 (s, 1H), 12.10 (s, 1H), 10.12 (s, 1H), 7.94 (s, 1H), 7.44 (s, 2H),7.29 (m, 4H), 3.54 (t, 2H, J=6.3 Hz), 2.34 (m, 2H), 1.83 (m, 2H), 1.62(m, 2H), 1.45 (m, 2H); ¹³C NMR (100 MHz, CD₃OD) δ 173.0, 148.0, 140.0,131.0, 129.6, 128.5, 128.0, 120.1, 116.1, 109.0, 36.6, 33.01, 32.5,27.6, 24.8; HRMS (ESI) calcd for C₁₅H₁₉BrN₄O (M⁺) 350.0742, found350.0738.

Cyclobutanecarboxylic acid [3-(2-amino-3H-imidazol-4-yl)phenyl]amidehydrochloride

In a similar manner, 0.050 g (0.140 mmol) of2-amino-5-[3-(cyclobutanecarbonylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester afforded 0.059 g (85%) of cyclobutanecarboxylicacid [3-(2-amino-3H-imidazol-4-yl)-phenyl]amide in its correspondinghydrochloride salt form as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ12.71 (s, 1H), 12.10 (s, 1H), 9.90 (s, 1H), 7.97 (s, 1H), 7.43 (s, 2H),7.34 (m, 4H), 3.26 (t, 1H, J=8.1 Hz), 2.17 (m, 4H), 1.94 (m, 2H), ¹³CNMR (100 MHz, CD₃OD) δ 176.4, 161.5, 150.0, 141.0, 130.8, 129.6, 121.4,121.3, 117.4, 110.3, 41.7, 26.2, 19.2; HRMS (ESI) calcd for C₁₄H₁₇N₄O(M⁺) 256.1324, found 256.1323.

5-(3-aminophenyl)-1H-imidazol-2-ylamine hydrochloride

In a similar manner, 0.045 g (0.165 mmol) of2-amino-5-(3-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.033 g (97%) of 5-(3-aminophenyl)-1H-imidazol-2-ylamine in itscorresponding hydrochloride salt form as a yellow amorphous solid: ¹HNMR (300 MHz, DMSO-d₆) δ 13.02 (s, 1H), 12.23 (s, 1H), 7.40-7.51 (m,8H), 7.17 (d, 1H, J=7.2 Hz); ¹³C NMR (100 MHz, CD₃OD) δ 149.5, 133.2,132.3, 131.3, 127.5, 126.2, 124.0, 120.4, 111.9; HRMS (ESI) calcd forC₉H₁₀N₄ (M⁺) 174.0906, found 174.0902.

Benzo[1,3]dioxole-5-carboxylic acid[3-(2-amino-3H-imidazol-4-yl)phenyl]amide hydrochloride

In a similar manner, 0.049 g (0.116 mmol) of2-amino-5-{3-[(benzo[1,3]dioxole-5-carbonyl)-amino]phenyl}imidazole-1-carboxylicacid tert-butyl ester afforded 0.040 g (97%) ofbenzo[1,3]dioxole-5-carboxylic acid[3-(2-amino-3H-imidazol-4-yl)phenyl]amide in its correspondinghydrochloride salt form as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ12.85 (s, 1H), 12.18 (s, 1H), 10.23 (s, 1H), 8.09 (s, 1H), 7.31-7.63 (m,8H), 7.07 (d, 1H, J=8.1 Hz), 6.14 (s, 2H); ¹³C NMR (100 MHz, CD₃OD) δ164.6, 150.2, 147.9, 147.4, 139.3, 129.2, 126.5, 122.9, 120.5, 119.8,116.6, 109.5, 108.0, 107.7, 101.9; HRMS (ESI) calcd for C₁₇H₁₄N₄O₃ (M⁺)322.1066, found 322.1061.

thiophene-2-carboxylic acid [3-(2-amino-3H-imidazol-4-yl)phenyl]amidehydrochloride

In a similar manner, 0.025 g (0.065 mmol) of2-amino-5-{3-[(thiophene-2-carbonyl)amino]phenyl}imidazole-1-carboxylicacid tert-butyl ester afforded 0.020 g (95%) of thiophene-2-carboxylicacid [3-(2-amino-3H-imidazol-4-yl)phenyl]amide in its correspondinghydrochloride salt form as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ12.82 (s, 1H), 12.16 (s, 1H), 10.44 (s, 1H), 8.17 (m, 1H), 8.06 (s, 1H),7.88 (m, 1H), 7.57 (m, 1H), 7.42 (m, 4H), 7.33 (s, 1H), 7.24 (m, 1H);¹³C NMR (100 MHz, CD₃OD) δ 163.0, 149.2, 140.7, 140.6, 133.0, 130.9,130.5, 129.7, 129.2, 129.0, 122.3, 121.8, 118.5, 110.4; HRMS (ESI) calcdfor C₁₄H₁₂N₄OS (M⁺) 284.0732, found 284.0729.

Furan-2-carboxylic acid [3-(2-amino-3H-imidazol-4-yl)phenyl]amidehydrochloride

In a similar manner, 0.031 g (0.084 mmol) of2-amino-5-{3-[(furan-2-carbonyl)amino]phenyl}imidazole-1-carboxylic acidtert-butyl ester afforded 0.025 g (99%) of furan-2-carboxylic acid[3-(2-amino-3H-imidazol-4-yl)phenyl]amide in its correspondinghydrochloride salt form as a colorless film: ¹H NMR (300 MHz, DMSO-d₆) δ12.88 (s, 1H), 12.21 (s, 1H), 10.33 (s, 1H), 8.00 (m, 1H), 7.60 (m, 1H),7.52 (br s, 2H), 7.39 (m, 4H), 7.30 (s, 1H), 6.72 (m, 1H); ¹³C NMR (100MHz, CD₃OD) δ 159.1, 149.5, 148.9, 147.0, 140.1, 130.9, 129.6, 128.9,122.3, 121.8, 118.3, 116.5, 113.4, 110.3; HRMS (ESI) calcd forC₁₄H₁₂N₄O₂ (M⁺) 268.0960, found 268.0956.

3-(2-amino-3H-imidazol-4-yl)phenyl-3-cyclopentylpropionamidehydrochloride

In a similar manner, 0.040 g (0.100 mmol) of2-amino-5-[3-(3-cyclopentylpropionylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester afforded 0.033 g (99%) of3-(2-amino-3H-imidazol-4-yl)phenyl-3-cyclopentylpropionamide in itscorresponding hydrochloride salt form as a tan amorphous solid: ¹H NMR(300 MHz, DMSO-d₆) δ 12.87 (s, 1H), 12.21 (s, 1H), 10.13 (s, 1H), 7.93(s, 1H), 7.45 (m, 3H), 7.25-7.38 (m, 3H), 2.34 (t, 2H, J=7.2 Hz), 1.73(m, 3H), 1.46-1.64 (m, 6H), 1.10 (m, 2H); ¹³C NMR (100 MHz, CD₃OD) δ175.2, 149.4, 140.9, 130.8, 129.6, 129.0, 121.3, 117.3, 110.3, 41.2,37.5, 33.7, 33.3, 26.2; HRMS (ESI) calcd for C₁₇H₂₂N₄O (M⁺) 298.1794,found 298.1789.

3-(2-amino-3H-imidazol-4-yl)phenyl-4-pentylbenzamide hydrochloride

In a similar manner, 0.050 g (0.111 mmol) of2-amino-5-[3-(4-pentylbenzoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester afforded 0.042 g (98%) of3-(2-amino-3H-imidazol-4-yl)phenyl-4-pentylbenzamide in itscorresponding hydrochloride salt form as a tan amorphous solid: ¹H NMR(400 MHz, DMSO-d₆) δ 12.74 (s, 1H), 12.10 (s, 1H), 10.30 (s, 1H), 8.09(s, 1H), 7.88 (d, 2H, J=8.0 Hz), 7.57 (m, 1H), 7.30-7.45 (m, 7H), 2.64(t, 2H, J=7.2 Hz), 1.60 (m, 2H), 1.29 (m, 4H), 0.87 (t, 3H, J=6.6 Hz);¹³C NMR (100 MHz, CD₃OD) δ 169.5, 149.3, 149.0, 141.3, 133.8, 132.4,131.2, 130.2, 130.0, 129.3, 122.8, 122.1, 118.9, 110.7, 37.2, 33.1,32.6, 24.1, 14.8; HRMS (ESI) calcd for C₂₁H₂₄N₄O (M⁺) 348.1950, found348.1944.

Adamantane-1-carboxylic acid [3-(2-amino-3H-imidazol-4-yl)phenyl]amidehydrochloride

In a similar manner, 0.060 g (0.137 mmol) of5-{3-[(adamantane-1-carbonyl)amino]phenyl}-2-aminoimidazole-1-carboxylicacid tert-butyl ester afforded 0.050 g (98%) of adamantane-1-carboxylicacid [3-(2-amino-3H-imidazol-4-yl)phenyl]amide in its correspondinghydrochloride salt form as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ12.80 (s, 1H), 12.17 (s, 1H), 9.29 (s, 1H), 7.98 (s, 1H), 7.49 (m, 3H),7.32 (m, 3H), 2.02 (br s, 3H), 1.92 (br s, 6H), 1.71 (br s, 6H); ¹³C NMR(100 MHz, CD₃OD) δ 179.7, 149.5, 140.7, 130.7, 129.5, 129.1, 122.8,121.6, 119.0, 110.3, 42.8, 40.1, 37.7, 29.8; HRMS (ESI) calcd forC₂₀H₂₅N₄O (M⁺) 336.1950, found 336.1945.

2-ethylhexanoic acid 3-(2-amino-3H-imidazol-4-yl)phenylamidehydrochloride

In a similar manner, 0.052 g (0.130 mmol) of2-amino-5-[3-(2-ethylhexanoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester afforded 0.042 g (99%) of 2-ethylhexanoic acid3-(2-amino-3H-imidazol-4-yl)phenylamide in its correspondinghydrochloride salt form as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ12.79 (s, 1H), 12.18 (s, 1H), 10.06 (s, 1H), 7.99 (s, 1H), 7.43 (m, 3H),7.28 (m, 3H), 2.30 (m, 1H), 1.27-1.58 (m, 8H), 0.85 (m, 6H); ¹³C NMR(100 MHz, CD₃OD) δ 177.9, 149.5, 140.7, 130.9, 129.7, 129.0, 121.5,117.6, 110.3, 95.9, 50.8, 33.9, 31.1, 27.5, 23.9, 14.5, 12.5; HRMS (ESI)calcd for C₁₇H₂₄N₄O (M⁺) 300.1950, found 300.1946.

3-(2-amino-3H-imidazol-4-yl)phenyl]-3,5-difluorobenzamide hydrochloride

In a similar manner, 0.041 g (0.098 mmol) of2-amino-5-[3-(3,5-difluorobenzoylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester afforded 0.034 g (98%) of3-(2-amino-3H-imidazol-4-yl)phenyl]-3,5-difluorobenzamide in itscorresponding hydrochloride salt form as a white solid: ¹H NMR (300 MHz,DMSO-d₆) δ 12.79 (s, 1H), 12.13 (s, 1H), 10.57 (s, 1H), 8.09 (s, 1H),7.72 (d, 2H, J=6.0 Hz), 7.53 (m, 2H), 7.44 (m, 4H), 7.32 (s, 1H); ¹³CNMR (100 MHz, CD₃OD) δ 166.5, 149.5, 148.1, 140.5, 130.9, 129.7, 128.9,122.4, 122.1, 118.5, 112.2, 112.0, 110.5, 108.2; HRMS (ESI) calcd forC₁₆H₁₂F₂N₄O (M⁺) 314.0979, found 314.0974.

Cyclohexanecarboxylic acid [3-(2-amino-3H-imidazol-4-yl)phenyl]amidehydrochloride

In a similar manner, 0.050 g (0.130 mmol) of2-amino-5-[3-(cyclohexanecarbonylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester afforded 0.038 g (92%) of cyclohexanecarboxylicacid [3-(2-amino-3H-imidazol-4-yl)phenyl]amide in its correspondinghydrochloride salt form as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ12.79 (s, 1H), 12.20 (s, 1H), 9.93 (s, 1H), 8.00 (s, 1H), 7.47 (s, 2H),7.35 (m, 2H), 7.28 (m, 2H), 2.34 (m 1H), 1.80 (m, 2H), 1.74 (m, 2H),1.40 (m, 2H), 1.94 (m, 2H), 1.23 (m, 2H); ¹³C NMR (100 MHz, CD₃OD) δ179.1, 150.0, 142.0, 130.8, 129.5, 129.0, 121.3, 121.2, 117.4, 110.2,48.8, 30.8, 27.0, 26.9; HRMS (ESI) calcd for C₁₆H₂₀N₄O (M⁺) 284.1637,found 284.1632.

Heptanoic acid [3-(2-amino-3H-imidazol-4-yl)phenyl]amide hydrochloride

In a similar manner, 0.056 g (0.110 mmol) of2-amino-5-(3-heptanoylaminophenyl)imidazole-1-carboxylic acid tert-butylester afforded 0.046 g (98%) of heptanoic acid[3-(2-amino-3H-imidazol-4-yl)phenyl]amide in its correspondinghydrochloride salt form as an off-white solid: ¹H NMR (300 MHz, DMSO-d₆)δ 12.72 (s, 1H), 12.10 (s, 1H), 10.05 (s, 1H), 7.95 (s, 1H), 7.43 (m,2H), 7.30 (m, 3H), 7.28 (s, 1H), 2.32 (t, 2H, J=7.5 Hz), 1.59 (m, 2H),1.28 (m, 6H), 0.87 (m, 3H); ¹³C NMR (100 MHz, CD₃OD) δ 175.1, 159.0,149.0, 140.9, 130.8, 129.6, 129.0, 121.3, 117.3, 110.3, 38.2, 32.9,30.1, 27.0, 23.7, 14.5; HRMS (ESI) calcd for C₁₆H₂₂N₄O (M⁺) 286.1794,found 286.1790.

3-(2-amino-3H-imidazol-4-yl)phenyl]-4-hexylbenzamide hydrochloride

In a similar manner, 0.070 g (0.151 mmol) of2-amino-5-[3-(4-hexylbenzoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester afforded 0.059 g (98%) of3-(2-amino-3H-imidazol-4-yl)phenyl]-4-hexylbenzamide in itscorresponding hydrochloride salt form as a white solid: ¹H NMR (300 MHz,DMSO-d₆) δ 12.77 (s, 1H), 12.12 (s, 1H), 10.32 (s, 1H), 8.02 (s, 1H),7.90 (d, 2H, J=8.1 Hz), 7.46 (s, 2H), 7.29-7.43 (m, 6H), 2.67 (t, 2H,J=7.5 Hz), 1.60 (m, 2H), 1.29 (m, 6H), 0.86 (m, 3H); ¹³C NMR (100 MHz,CD₃OD) δ 167.7, 147.6, 139.6, 132.1, 129.7, 129.5, 128.6, 128.4, 127.9,127.7, 121.1, 120.4, 117.1, 109.0, 35.6, 31.7, 31.2, 28.9, 22.5, 13.3;HRMS (ESI) calcd for C₂₂H₂₆N₄O (M⁺) 326.2107, found 362.2101.

2-amino-5-[2-(cyclopropanecarbonylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(2-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.076 g (82%) of2-amino-5-[2-(cyclopropanecarbonylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester as an off-white solid: ¹H NMR (300 MHz, DMSO-d₆) δ12.37 (s, 1H), 8.31 (d, 1H, J=8.1 Hz), 7.70 (dd, 1H, J=1.2, 7.8 Hz),7.38 (s, 1H), 7.18 (m, 1H), 7.01 (t, 1H, J=7.2 Hz), 6.93 (br s, 2H),1.88 (m, 1H), 1.59 (s, 9H), 0.83 (m, 4H); ¹³C NMR (100 MHz, CD₃OD) δ171.3, 149.6, 148.7, 136.5, 135.8, 127.6, 126.5, 122.9, 120.7, 120.0,107.6, 85.2, 27.5, 15.6, 7.3; HRMS (ESI) calcd for C₁₈H₂₂N₄O₃ (M⁺)342.1692, found 342.1687.

2-amino-5-(2-decanoylaminophenyl)imidazole-1-carboxylic acid tert-butylester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(2-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.084 g (72%) of2-amino-5-(2-decanoylaminophenyl)imidazole-1-carboxylic acid tert-butylester as a tan oil: ¹H NMR (400 MHz, DMSO-d₆) δ 11.89 (s, 1H), 8.30 (d,1H, J=8.0 Hz), 7.70 (dd, 1H, J=1.2, 8.0 Hz), 7.34 (s, 1H), 7.19 (m, 1H),7.02 (t, 1H, J=6.8 Hz), 6.88 (br s, 2H), 2.37 (t, 2H, J=7.2 Hz), 1.59(m, 11H), 1.25 (m, 12H), 0.83 (t, 3H, J=6.4 Hz); ¹³C NMR (100 MHz,DMSO-d₆) δ 171.0, 149.6, 148.7, 136.3, 135.8, 127.5, 126.6, 123.0,120.9, 120.5, 107.5, 85.1, 37.4, 31.3, 28.9, 28.8, 28.6, 28.5, 27.5,25.2, 22.1, 14.0; HRMS (ESI) calcd for C₂₄H₃₆N₄O₃ (M⁺) 428.2787, found428.2782.

2-amino-5-[2-(cyclohexanecarbonylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(2-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.078 g (74%) of2-amino-5-[2-(cyclohexanecarbonylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a white solid: ¹H NMR (400 MHz, DMSO-d₆) δ11.65 (s, 1H), 8.26 (d, 1H, J=7.6 Hz), 7.70 (dd, 1H, J=1.2, 7.6 Hz),7.32 (s, 1H), 7.18 (m, 1H), 7.02 (t, 1H, J=6.8 Hz), 6.86 (br s, 2H),2.34 (m, 1H), 1.83 (m, 2H), 1.74 (m, 2H), 1.59 (m, 11H), 1.31-1.48 (m,3H), 1.20 (m, 1H); ¹³C NMR (100 MHz, CD₃OD) δ 177.5, 150.8, 137.5,137.2, 129.0, 128.3, 128.1, 125.4, 124.2, 123.7, 109.1, 86.9, 48.1,30.9, 28.3, 27.1, 26.9; HRMS (ESI) calcd for C₂₁H₂₈N₄O₃ (M⁺) 384.2161,found 384.2157.

2-amino-5-[2-(4-pentylbenzoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester

In a similar manner, 0.075 g (0.273 mmol) of2-amino-5-(2-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.089 g (73%) of2-amino-5-[2-(4-pentylbenzoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester as a white foam: ¹H NMR (400 MHz, DMSO-d₆) δ 12.41 (s,1H), 8.52 (d, 1H, J=7.6 Hz), 7.99 (d, 2H, J=8.4 Hz), 7.71 (dd, 1H,J=1.6, 8.0 Hz), 7.41 (d, 2H, J=8.4 Hz), 7.37 (s, 1H), 7.29 (m, 1H), 7.10(m, 1H), 6.93 (br s, 2H), 2.66 (t, 2H, J=7.2 Hz), 1.61 (m, 2H), 1.57 (s,9H), 1.31 (m, 4H), 0.86 (t, 3H, J=6.8 Hz); ¹³C NMR (100 MHz, CD₃OD) δ169.1, 150.8, 148.8, 137.8, 137.6, 133.7, 130.1, 130.0, 129.1, 128.8,128.2, 125.3, 123.8, 123.1, 109.1, 86.9, 36.9, 32.8, 32.3, 28.3, 23.7,14.5; HRMS (ESI) calcd for C₂₆H₃₂N₄O₃ (M⁺) 448.2474, found 448.2466.

2-amino-5-[2-(6-bromohexanoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester

In a similar manner, 0.071 g (0.258 mmol) of2-amino-5-(2-aminophenyl)imidazole-1-carboxylic acid tert-butyl esterafforded 0.085 g (75%) of2-amino-5-[2-(6-bromohexanoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester as an off-white solid: ¹H NMR (300 MHz, DMSO-d₆) δ11.92 (s, 1H), 8.31 (d, 1H, J=7.5 Hz), 7.69 (dd, 1H, J=1.2, 8.1 Hz),7.35 (s, 1H), 7.20 (m, 1H), 7.02 (m, 1H), 6.89 (br s, 2H), 3.52 (t, 2H,J=6.3 Hz), 2.39 (t, 2H, J=7.5 Hz), 1.84 (m, 2H), 1.64 (m, 2H), 1.59 (s,9H), 1.43 (m, 2H); ¹³C NMR (100 MHz, CDCl₃) δ 171.4, 149.5, 149.3,137.3, 137.0, 128.6, 126.6, 123.2, 121.2, 120.1, 107.8, 86.1, 38.4,33.9, 32.7, 28.1, 28.0, 24.9; HRMS (ESI) calcd for C₂₀H₂₇BrN₄O₃ (M⁺)450.1267, found 450.1260.

2-(2-amino-3H-imidazol-4-yl)phenyl-4-pentyl-benzamide hydrochloride

In a similar manner, 0.044 g (0.098 mmol) of2-amino-5-[2-(4-pentylbenzoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester afforded 0.037 g (99%) of2-(2-amino-3H-imidazol-4-yl)phenyl-4-pentyl-benzamide in itscorresponding hydrochloride salt form as a white solid: ¹H NMR (300 MHz,DMSO-d₆) δ 12.69 (s, 1H), 11.93 (s, 1H), 10.02 (s, 1H), 7.88 (d, 2H,J=8.1 Hz), 7.53 (m, 2H), 7.46 (m, 2H), 7.40 (m, 1H), 7.33 (d, 2H, J=8.1Hz), 6.96 (s, 1H), 2.64 (t, 2H, J=7.2 Hz), 1.60 (m, 2H), 1.29 (m, 4H),0.87 (t, 3H, J=6.6 Hz); ¹³C NMR (100 MHz, CD₃OD) δ 169.5, 149.2, 149.0,136.3, 132.7, 130.7, 130.0, 129.9, 129.4, 129.0, 128.5, 126.3, 126.1,112.0, 36.9, 32.7, 32.3, 23.7, 14.5; HRMS (ESI) calcd for C₂₁H₂₄N₄O (M⁺)348.1950, found 348.1945.

decanoic acid 2-(2-amino-3H-imidazol-4-yl)phenyl amide hydrochloride

In a similar manner, 0.046 g (0.107 mmol) of2-amino-5-(2-decanoylaminophenyl)imidazole-1-carboxylic acid tert-butylester afforded 0.039 g (99%) of decanoic acid2-(2-amino-3H-imidazol-4-yl)phenyl amide in its correspondinghydrochloride salt form as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ12.58 (s, 1H), 12.10 (s, 1H), 9.47 (s, 1H), 7.46 (m, 4H), 7.36 (m, 1H),7.27 (t, 1H, J=7.2 Hz), 7.03 (s, 1H), 2.30 (t, 2H, J=6.9 Hz), 1.55 (m,2H), 1.25 (br s, 12H), 0.86 (t, 3H, J=6.6 Hz); ¹³C NMR (100 MHz, CD₃OD)δ 175.7, 149.0, 136.2, 130.7, 130.1, 128.6, 128.0, 125.8, 125.3, 112.2,37.5, 33.2, 30.7, 30.6, 30.5, 26.9, 23.9, 14.6; HRMS (ESI) calcd forC₁₉H₂₈N₄O (M⁺) 328.2263, found 328.2260.

6-bromohexanoic acid-2-(2-amino-3H-imidazol-4-yl)phenyl amidehydrochloride

In a similar manner, 0.038 g (0.084 mmol) of2-amino-5-[2-(6-bromohexanoylamino)phenyl]imidazole-1-carboxylic acidtert-butyl ester afforded 0.032 g (97%) of 6-bromohexanoicacid-2-(2-amino-3H-imidazol-4-yl)phenyl amide in its correspondinghydrochloride salt form as a tan solid: ¹H NMR (400 MHz, DMSO-d₆) δ12.64 (s, 1H), 12.19 (s, 1H), 9.46 (s, 1H), 7.53 (m, 3H), 7.44 (d, 1H,J=7.2 Hz), 7.36 (t, 1H, J=6.8 Hz), 7.27 (t, 1H, J=7.2 Hz), 7.03 (s, 1H),3.54 (t, 2H, J=7.2 Hz), 2.32 (t, 2H, J=6.9 Hz), 1.82 (tt, 2H, J=6.9,14.0 Hz), 1.59 (tt, 2H, J=6.8, 14.4 Hz), 1.41 (tt, 2H, J=8.0, 14.8 Hz);¹³C NMR (100 MHz, CD₃OD) δ 175.3, 149.5, 136.2, 130.7, 130.2, 128.6,128.0, 125.8, 125.3, 112.2, 37.2, 34.3, 33.7, 29.0, 25.9; HRMS (ESI)calcd for C₁₅H₁₉BrN₄O (M⁺) 350.0742, found 350.0739.

Cyclopropanecarboxylic acid 2-(2-amino-3H-imidazol-4-yl)phenyl amidehydrochloride

In a similar manner, 0.046 g (0.134 mmol) of2-amino-5-[2-(cyclopropanecarbonylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester afforded 0.036 g (97%) of cyclopropanecarboxylicacid 2-(2-amino-3H-imidazol-4-yl)phenyl amide in its correspondinghydrochloride salt form as a tan solid: ¹H NMR (300 MHz, DMSO-d₆) δ12.69 (s, 1H), 12.17 (s, 1H), 9.82 (s, 1H), 7.51 (m, 4H), 7.35 (m, 1H),7.28 (t, 1H, J=6.3 Hz), 7.03 (s, 1H), 1.88 (m, 1H), 0.76 (d, 4H, J=5.7Hz); ¹³C NMR (100 MHz, CD₃OD) δ 176.0, 149.1, 136.3, 130.7, 129.9,128.5, 127.9, 125.6, 125.1, 112.3, 15.4, 8.2; HRMS (ESI) calcd forC₁₃H₁₄N₄O (M⁺) 242.1168, found 242.1164.

Cyclohexanecarboxylic acid 3-(2-amino-3H-imidazol-4-yl)phenyl amidehydrochloride

In a similar manner, 0.041 g (0.106 mmol) of2-amino-5-[2-(cyclohexanecarbonylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester afforded 0.032 g (94%) of cyclohexanecarboxylicacid 3-(2-amino-3H-imidazol-4-yl)phenyl amide in its correspondinghydrochloride salt form as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ12.58 (s, 1H), 12.05 (s, 1H), 9.41 (s, 1H), 7.45 (m, 4H), 7.35 (m, 1H),7.28 (t, 1H, J=7.8 Hz), 6.99 (s, 1H), 2.36 (m, 1H), 1.63-1.83 (m, 4H),1.20-1.41 (m, 6H); ¹³C NMR (100 MHz, CD₃OD) δ 178.4, 148.9, 136.3,132.5, 130.8, 130.2, 128.7, 128.1, 125.7, 112.2, 46.6, 30.6, 27.0, 26.9;HRMS (ESI) calcd for C₁₆H₂₀N₄O (M⁺) 284.1637, found 284.1635.

Cyclobutanecarboxylic acid [2-(2-amino-3H-imidazol-4-yl)phenyl]amidehydrochloride

In a similar manner, 0.044 g (0.122 mmol) of2-amino-5-[2-(cyclobutanecarbonylamino)phenyl]imidazole-1-carboxylicacid tert-butyl ester afforded 0.034 g (98%) of cyclobutanecarboxylicacid [2-(2-amino-3H-imidazol-4-yl)phenyl]amide in its correspondinghydrochloride salt form as a yellow solid: ¹H NMR (300 MHz, DMSO-d₆) δ12.47 (s, 1H), 12.00 (s, 1H), 9.30 (s, 1H), 7.53 (d, 1H, J=6.0), 7.45(s, 2H), 7.35 (t, 1H, J=6.4 Hz), 7.29 (t, 1H, J=7.2 Hz), 6.96 (s, 1H),3.25 (t, 1H, J=6.3 Hz), 2.04 (m, 2H), 2.50 (m, 2H), 1.48 (m, 2H); ¹³CNMR (100 MHz, CD₃OD) δ 176.9, 148.0, 136.2, 130.7, 130.0, 128.6, 128.0,125.9, 125.3, 112.2, 41.2, 26.2, 19.2; HRMS (ESI) calcd for C₁₄H₁₇N₄O(M⁺) 256.1324, found 256.1319.

Example 5: Synthesis of 2-Aminoimidazole Libraries for Anti-BiofilmScreening

The following compounds have been synthesized.

2-amino-5-(4-azidophenyl)imidazole-1-carboxylic acid tert-butyl ester(0.229 g, 0.762 mmol) was dissolved in anhydrous THF (2.5 mL) and cooledto 0° C. NaHMDS (1M in THF) (1.53 mL, 1.53 mmol) was added to thereaction dropwise. Boc-anhydride (0.174 g, 0.800 mmol) was dissolved inanhydrous THF (2 mL) and added to the reaction. The reaction was allowedto stir for 15 min at 0° C. then allowed to stir at warm to roomtemperature for 20 min. After the reaction was quenched with sat. NH₄Cland diluted with EtOAc (50 mL). The organic layer was then extractedwith brine (3×20 mL) before being dried (NaSO₄), filtered, andevaporated to dryness. The crude product was purified by columnchromatography (10-30% EtOAc/Hexanes) to afford 0.230 g (75%) of thetarget compound5-(4-azidophenyl)-2-tert-butoxycarbonylaminoimidazole-1-carboxylic acidtert-butyl ester as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ 9.54 (s,1H), 7.88 (s, 1H), 8.5 (d, 2H, J=8.7 Hz), 7.14 (d, 2H, J=8.7 Hz), 1.57(s, 9H), 1.44 (s, 9H); ¹³C NMR (75 MHz, DMSO-d₆) δ 152.8, 146.7, 139.6,138.1, 136.2, 129.7, 126.2, 119.3, 112.5, 85.2, 80.0, 70.8, 27.9, 27.3;HRMS (ESI) calcd for C₁₉H₂₄N₆O₄ (M⁺) 400.1857, found 400.1859.

5-(4-azidophenyl)-2-tert-butoxycarbonylaminoimidazole-1-carboxylic acidtert-butyl ester (0.080 g, 0.200 mmol), 1-ethynyl-3,5-difluorobenzene(0.035 g, 0.26 mmol), CuSO₄.5H₂O (0.097 g, 0.040 mmol), and sodiumascorbate (0.004 g, 0.020 mmol) were all charged in a reaction vesseland dissolved in a 1:1:1 mixture of H₂O (0.75 mL), EtOH 0.75 mL), andCH₂Cl₂ (0.75 mL). The reaction was stirred at ambient temperature for 16h. After the reaction was quenched with sat. NaHCO₃ and diluted withEtOAc (50 mL). The organic layer was washed with sat. NaHCO₃ (3×20 mL)before being dried (Na₂SO₄), filtered and evaporated to dryness. Thecrude product was purified by column chromatography (10%-30%EtOAc/Hexanes) to afford 0.068 g (44%) of2-tert-butoxycarbonylamino-5-{4-[4-(4-pentylphenyl)-[1,2,3]triazol-1-yl]phenyl}imidazole-1-carboxylicacid tert-butyl ester as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ 9.60(s, 1H), 9.29 (s, 1H), 8.01 (m, 3H), 7.98 (d, 2H, J=6.0 Hz), 7.85 (d,2H, J=5.6 Hz), 7.32 (d, 2H, J=5.4 Hz), 2.62 (t, 2H, J=5.1 Hz), 1.59 (s,9H), 1.47 (m, 2H), 1.45 (s, 9H), 1.30 (m, 4H), 0.87 (m, 3H); ¹³C NMR (75MHz, DMSO-d₆) δ 152.8, 147.2, 142.3, 139.9, 135.5, 128.9, 127.8, 127.6,125.9, 120.1, 118.9, 113.5, 85.5, 80.0, 38.7, 31.3, 30.8, 27.9, 27.3,21.9, 13.9; HRMS (ESI) calcd for C₃₂H₄₀N₆O₄ (M⁺) 572.3111, found572.3113.

In a similar manner, 0.051 g (0.127 mmol)5-(4-azidophenyl)-2-tert-butoxycarbonylaminoimidazole-1-carboxylic acidtert-butyl ester afforded 0.044 g (68%) of compound2-tert-butoxycarbonylamino-5-{4-[4-(3-chloropropyl)-[1,2,3]triazol-1-yl]phenyl}imidazole-1-carboxylicacid tert-butyl ester as a yellow solid: ¹H NMR (300 MHz, DMSO-d₆) δ9.18 (s, 1H), 7.95 (d, 2H, J=4.8 Hz), 7.78 (s, 1H), 7.68 (m, 2H), 7.24(s, 1H), 3.60 (t, 2H, J=6.3 Hz), 2.96 (t, 2H, J=7.5 Hz), 2.23 (m, 2H);¹³C NMR (75 MHz, DMSO-d₆) δ 150.1, 149.4, 147.3, 142.8, 137.9, 136.4,133.3, 126.9, 120.5, 119.4, 108.5, 87.2, 82.2, 44.3, 32.0, 28.4, 28.2,22.9; HRMS (ESI) calcd for C₂₄H₃₂N₆O₄Cl (M⁺) 502.2095, found 502.2094.

In a similar manner, 0.074 g (0.185 mmol) of5-(4-azidophenyl)-2-tert-butoxycarbonylaminoimidazole-1-carboxylic acidtert-butyl ester afforded 0.084 g (35%) of compound2-tert-butoxycarbonylamino-5-[4-(4-hydroxymethyl-[1,2,3]triazol-1-yl)phenyl]imidazole-1-carboxylicacid tert-butyl ester as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ 9.59(s, 1H), 8.71 (s, 1H), 7.99 (m, 3H), 7.92 (d, 2H, J=9.0 Hz), 4.61 (d,2H, J=3.0 Hz), 1.59 (s, 9H), 1.48 (s, 9H); ¹³C NMR (75 MHz, DMSO-d₆) δ153.0, 149.5, 139.4, 135.9, 134.4, 132.8, 125.9, 125.0, 120.8, 119.1,113.5, 85.4, 80.2, 54.9, 27.9, 27.7; HRMS (ESI) calcd for C₂₂H₂₈N₆O₅(M⁺) 456.2121, found 456.2121.

In a similar manner, 0.080 g (0.199 mmol) of5-(4-azidophenyl)-2-tert-butoxycarbonylaminoimidazole-1-carboxylic acidtert-butyl ester afforded 0.108 g (56%) of compound2-tert-butoxycarbonylamino-5-{4-[4-(3,5-difluorophenyl)-[1,2,3]triazol-1-yl]phenyl}imidazole-1-carboxylicacid tert-butyl ester as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ 9.61(s, 1H), 9.48 (s, 1H), 8.10 (m, 3H), 7.95 (d, 2H, J=8.7 Hz), 7.67 (m,2H), 7.30 (m, 1H), 1.59 (s, 9H), 1.45 (s, 9H); ¹³C NMR (75 MHz, DMSO-d₆)δ 164.4, 161.2, 152.8, 146.7, 145.3, 139.9, 135.9, 135.2, 133.3, 126.0,120.9, 120.2, 113.6, 108.4, 108.1, 85.4, 80.1, 27.9, 27.3; HRMS (ESI)calcd for C₂₇H₂₈N₆O₄F₂ (M⁺) 538.2140, found 538.2143.

In a similar manner, 0.070 g (0.175 mmol)5-(4-azidophenyl)-2-tert-butoxycarbonylaminoimidazole-1-carboxylic acidtert-butyl ester afforded 0.060 g (73%) of compound2-tert-butoxycarbonylamino-5-[4-(4-cyclopropyl-[1,2,3]triazol-1-yl)-phenyl]imidazole-1-carboxylicacidtert-butyl ester as a white solid: ¹H NMR (300 MHz, DMSO-d₆) δ 9.60 (s,1H), 8.56 (s, 1H), 8.10 (m, 3H), 7.87 (d, 2H, J=8.5 Hz), 2.00 (m, 1H),1.50 (s, 9H), 1.40 (s, 9H) 0.98 (m, 2H), 0.810 (m, 2H); ¹³C NMR (75 MHz,DMSO-d₆) δ 153.5, 152.7, 150.7, 134.8, 124.3, 119.4, 118.9, 80.75, 68.0,48.9, 31.5, 28.3, 28.0, 8.2, 6.9; HRMS (ESI) calcd for C₂₄H₃₀N₆O₄ (M⁺)466.2329, found 466.2326.

2-tert-butoxycarbonylamino-5-[4-(4-cyclopropyl-[1,2,3]triazol-1-yl)phenyl]imidazole-1-carboxylicacidtert-butyl ester (0.047 g, 0.089 mmol) was dissolved in anhydrousdichloromethane (5 mL) and cooled to 0° C. Trifluoroacetic acid (0.5 mL)was added drop-wise while the reaction continued to stir at 0° C. Uponcompletion, the reaction was allowed to warm to room temperature overthe course of 15 h. Then the reaction was evaporated down. The crude TFAsalt was the dissolved in dichloromethane (2 mL) and a 2M solution ofHCl in diethyl ether (0.100 mL) was added. The solution was againconcentrated down under vacuum to afford 0.033 g (98%) of desiredproduct5-[4-(4-cyclopropyl-[1,2,3]triazol-1-yl)phenyl]-1H-imidazol-2-ylamine inits corresponding hydrochloride salt form as an off-white solid: ¹H NMR(300 MHz, DMSO-d₆) δ 13.20 (s, 1H), 12.30 (s, 1H), 8.59 (s, 1H), 7.92(m, 4H), 7.52 (m, 3H), 2.01 (m, 1H), 0.98 (m, 2H), 0.80 (m, 2H); ¹³C NMR(100 MHz, DMSO-d₆) δ 151.4, 147.6, 135.6, 127.4, 125.3, 120.0, 118.5,110.2, 94.0, 7.6, 6.2; HRMS (ESI) calcd for C₁₄H₁₄N₆ (M⁺) 266.1280,found 266.1282.

In a similar manner, 0.047 g (0.088 mmol)2-tert-butoxycarbonylamino-5-{4-[4-(3,5-difluorophenyl)-[1,2,3]triazol-1-yl]-phenyl}imidazole-1-carboxylicacid tert-butyl ester afforded 0.033 g (98%) of compound5-{4-[4-(3,5-difluorophenyl)-[1,2,3]triazol-1-yl-phenyl}-1H-imidazol-2-ylaminein its corresponding hydrochloride salt form as a white solid: ¹H NMR(300 MHz, DMSO-d₆) δ 13.05 (s, 1H), 12.15 (s, 1H), 9.51 (s, 1H), 8.03(m, 4H), 7.66 (m, 2H), 7.57 (s, 3H), 7.28 (m, 1H); ¹³C NMR (75 MHz,DMSO-d₆) δ 161.2, 156.4, 147.8, 145.3, 135.4, 128.2, 125.5, 125.4,120.8, 120.6, 110.6, 108.4, 108.1; HRMS (ESI) calcd for C₁₇H₁₂N₆F₂ (M⁺)338.1092, found 338.1093.

In a similar manner, 0.051 g (0.090 mmol)2-tert-butoxycarbonylamino-5-{4-[4-(4-pentylphenyl)-[1,2,3]triazol-1-yl]-phenyl}imidazole-1-carboxylicacid tert-butyl ester afforded 0.036 g (98%) of compound5-{4-[4-(4-pentylphenyl)-[1,2,3]triazol-1-yl-phenyl}-1H-imidazol-2-ylaminein its corresponding hydrochloride salt form as a brown solid: ¹H NMR(300 MHz, DMSO-d₆) δ 13.12 (s, 1H), 12.27 (s, 1H), 9.31 (s, 1H), 8.06(d, 2H, J=9.0 Hz), 7.93 (d, 2H, J=8.4 Hz), 7.85 (d, 2H, J=8.1 Hz), 7.57(m, 3H), 7.32 (d, 2H, J=8.1 Hz), 2.62 (t, 2H, J=7.2 Hz), 1.60 (m, 2H),1.29 (m, 4H), 0.87 (t, 3H, J=6.6 Hz); ¹³C NMR (75 MHz, DMSO-d₆) δ 147.9,147.5, 142.6, 135.7, 128.8, 127.9, 127.6, 125.5, 125.4, 125.3, 120.2,119.0, 110.5, 35.6, 31.6, 31.2, 22.6, 14.6; HRMS (ESI) calcd forC₂₂H₂₄N₆ (M⁺) 372.2062, found 372.2062.

In a similar manner, 0.028 g (0.057 mmol)2-tert-butoxycarbonylamino-5-{4-(4-(3-chloropropyl)-[1,2,3]triazol-1-yl)phenyl}imidazole-1-carboxylicacid tert-butyl ester afforded 0.018 g (93%) of compound5-{4-[4-(3-chloropropyl-[1,2,3]triazol-1-yl]phenyl}-1H-imidazol-2-ylaminein its corresponding hydrochloride salt form as a brown solid: ¹H NMR(300 MHz, DMSO-d₆) δ 13.06 (s, 1H), 12.24 (s, 1H), 8.61 (s, 1H), 7.97(d, 2H, J=9.0 Hz), 7.88 (d, 2H, J=8.7 Hz), 7.54 (m, 3H), 3.74 (t, 2H,J=6.3 Hz), 2.86 (m, 2H, J=6.9 Hz), 2.13 (m, 2H); ¹³C NMR (100 MHz,DMSO-d₆) δ 149.8, 148.0, 137.6, 130.4, 127.7, 127.2, 123.5, 122.6,111.8, 45.0, 33.0 23.3; HRMS (ESI) calcd for C₁₄H₁₅N₆Cl (M⁺) 302.1050,found 302.1047.

In a similar manner, 0.020 g (0.045 mmol)2-tert-butoxycarbonylamino-5-[4-(4-hydroxymethyl-[1,2,3]triazol-1-yl)phenyl]imidazole-1-carboxylicacid tert-butyl ester afforded 0.014 g (98%) of compound{1-[4-(2-amino-3H-imidazol-4-yl)phenyl]-1H-[1,2,3]triazol-4-yl}methanolin its corresponding hydrochloride salt form as an off-white solid: ¹HNMR (300 MHz, CD₃OD) δ 8.52 (s, 1H), 7.85 (d, 2H, J=8.7 Hz), 7.70 (d,2H, J=8.7 Hz), 7.20 (s, 1H), 4.72 (m, 2H); ¹³C NMR (75 MHz, CD₃OD) δ178.5, 136.2, 129.0, 127.0, 126.6, 122.0, 121.5, 111.1, 71.0, 55.0; HRMS(ESI) calcd for C₁₂H₁₂N₆₀ (M⁺) 256.1073, found 256.1074.

Example 6: Activity Testing of Compounds Against Bacterial Strains

General Static Bacterial Biofilm Inhibition Assay Procedure for A.baumannii, P. aeruginosa and S. aureus:

Biofilm inhibition assays are performed by taking an overnight cultureof bacterial strain and subculturing it at an OD₆₀₀ of 0.10 into thenecessary growth liquid medium (LB for A. baumannii, LBNS for P.aeruginosa, and TSBG for S. aureus) for the strain. The compound beingtested is then added at a predetermined concentration and then aliquoted(100 μL) into the wells of a 96-well PVC microtiter plate (Wells notused for samples are filled with 100 μL of de-ionized water). Plates arethen wrapped in GLAD Press n' Seal® and incubated under stationaryconditions at 37° C. After 24 hours, the media is discarded from thewells and the plates are washed thoroughly with tap water. Plates arethen stained with 100 μL of 0.1% solution of crystal violet (CV) andthen incubated at an ambient temperature for 30 minutes. Sample platesare then washed with tap water again, and the remaining stain issolubilized with 200 μL of 95% ethanol. Biofilm inhibition osquantitated by measuring the OD₅₄₀ for each well by transferring 125 μLof the solubilized CV stain into a polystyrene microtiter dish foranalysis.

General Static Bacterial Biofilm Dispersion Assay Procedure for A.baumannii, P. aeruginosa and S. aureus:

Dispersion assays are performed by taking an overnight culture ofbacterial strain and subculturing it at an OD₆₀₀ of 0.10 into thenecessary growth liquid medium. The resulting bacterial suspension isaliquoted (100 μL) into the wells of a 96-well PVC microtiter plate.Plates are then wrapped in GLAD Press n' Seal® followed by an incubationunder stationary conditions at an ambient temperature. After 24 hours,the media is discarded from the wells and the plates are washedthoroughly with tap water. Predetermined concentrations of the compoundare then made in the same medium used to initially grow the biofilms andthen aliquoted (100 μL) into the wells of the 96-well PVC microtiterplate with the established biofilms. Plates are then wrapped in GLADPress n' Seal® and incubated under stationary conditions at 37° C. After24 hours, the media is discarded from the wells and the plates arewashed thoroughly with tap water. Plates are then stained with 100 μL of0.1% solution of crystal violet (CV) and then incubated at roomtemperature for 30 minutes. Plates are then washed with tap water againand the remaining stain was solubilized with 200 μL of 95% ethanol.Biofilm dispersion is quantitated by measuring the OD₅₄₀ for each wellby transferring 125 μL of the solubilized CV stain into a polystyrenemicrotiter dish for analysis.

General Planktonic Growth Curve Procedure:

Bacterial strains are grown in the absence or presence of the testcompound at the IC₅₀ value starting at an OD₆₀₀ of 0.1 in culture tubesin an incubator shaker at 37° C. at 200 rpm. The OD₆₀₀ is recorded at 1,3, 4, 5, 6 and 24 hrs.

General Colony Count Procedure for A. baumannii, P. aeruginosa and S.aeureus:

Colony counts are performed by incubating either bacterial strain in thepresence and absence of the test compound at 37° C. in culture tubesuntil the sample with the absence of the test compound reached an OD₆₀₀of 0.40 from a starting OD₆₀₀ of 0.10. This typically takes three tofour hours. Once the OD₆₀₀ of approximately 0.40 was observed, 100 μLare taken from each culture tube from which serial dilutions are made.Then, 10 μL are removed from each serial dilution and plated out on asquare gridded petri-dish followed by 16 hours of incubation at 37° C.period to grow countable colonies. Viable bacteria are quantifiedthrough employment of the track-dilution method (Jett et al., BioTechniques. 1997, 23, 648-650).

The foregoing is illustrative of the invention, and is not to beconstrued as limiting thereof. The invention is defined by the followingclaims, with equivalents to be included.

That which is claimed is:
 1. A compound of Formula (IV)(a):

wherein: R¹⁴ is selected from the group consisting of: heterocyclo,aryl, and heteroaryl; R^(14′), R¹² and R^(12′) are each independently Hor alkyl; and q=0 to 10, saturated or unsaturated; or a pharmaceuticallyacceptable salt or prodrug thereof.
 2. The compound of claim 1, whereinR¹⁴ is aryl, and said aryl is optionally substituted, or apharmaceutically acceptable salt or prodrug thereof.
 3. The compound ofclaim 1, wherein q=0.
 4. A composition comprising the compound of claim1 and a biocide.
 5. A composition comprising the compound of claim 1covalently coupled to a substrate.
 6. A biofilm preventing, removing orinhibiting coating composition, comprising: (a) a film-forming resin;(b) a solvent that disperses said resin; (c) an effective amount of thecompound of claim 1, wherein said effective amount of said compoundprevents or inhibits the growth of a biofilm thereon; and (d)optionally, at least one pigment.
 7. The compound of claim 1, whereinsaid compound is a compound of Formula (IV)(a)(3):

wherein R¹⁴ is selected from the group consisting of: heterocyclo, aryland heteroaryl; and q=0 to 10, saturated or unsaturated; or apharmaceutically acceptable salt or prodrug thereof.
 8. The compound ofclaim 1, wherein R¹⁴ is aryl, and said aryl is substituted 1-3 timeswith halo; or a pharmaceutically acceptable salt or prodrug thereof. 9.The compound of claim 8, wherein q=0; or a pharmaceutically acceptablesalt or prodrug thereof.
 10. The compound of claim 8, wherein saidcompound has the formula:

or a pharmaceutically acceptable salt or prodrug thereof.
 11. A medicaldevice comprising: (a) a medical device substrate; and (b) an effectiveamount of the compound of claim 1, either coating the substrate, orincorporated into the substrate, wherein said effective amount of saidcompound prevents or inhibits the growth of a biofilm thereon.
 12. Themedical device of claim 11, wherein said medical device substrate isselected from the group consisting of stents, fasteners, ports,catheters, scaffolds and grafts.
 13. The medical device of claim 11,wherein said compound is covalently coupled to said substrate.
 14. Amethod of controlling biofilm formation on a substrate comprising thestep of contacting the compound of claim 1 to said substrate in anamount effective to inhibit biofilm formation or clear a preformedbiofilm.
 15. The method of claim 14, wherein the substrate is selectedfrom the group consisting of a drainpipe, glaze ceramic, porcelain,glass, metal, wood, chrome, plastic, vinyl, and laminate.
 16. A compoundof Formula (II)(a):

wherein: R⁴ is selected from the group consisting of: H, hydroxy, acyl,alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl,alkoxy, amino, amide, sulfone, sulfoxide, oxo, nitro, and carboxy;R^(4′), R⁵ and R^(5′) are each independently H or alkyl; and n=0 to 10,saturated or unsaturated; or a pharmaceutically acceptable salt orprodrug thereof.
 17. The compound of claim 16, wherein R⁴ is selectedfrom the group consisting of: heterocyclo, aryl and heteroaryl.
 18. Thecompound of claim 16, wherein R⁴ is a group:

wherein: R⁷ is selected from the group consisting of: H, hydroxy, acyl,alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl,alkoxy, amino, amide, sulfone, sulfoxide, oxo, nitro, and carboxy; andm=0 to 10, saturated or unsaturated; or a pharmaceutically acceptablesalt or prodrug thereof.
 19. The compound of claim 16, wherein n=0. 20.The compound of claim 16, wherein said compound has the formula:

or a pharmaceutically acceptable salt or prodrug thereof.
 21. A biofilmpreventing, removing or inhibiting coating composition, comprising: (a)a film-forming resin; (b) a solvent that disperses said resin; (c) aneffective amount of the compound of claim 16, wherein said effectiveamount of said compound prevents or inhibits the growth of a biofilmthereon; and (d) optionally, at least one pigment.
 22. A medical devicecomprising: (a) a medical device substrate; and (b) an effective amountof the compound of claim 16, either coating the substrate, orincorporated into the substrate, wherein said effective amount of saidcompound prevents or inhibits the growth of a biofilm thereon.
 23. Themedical device of claim 22, wherein said medical device substrate isselected from the group consisting of stents, fasteners, ports,catheters, scaffolds and grafts.
 24. The medical device of claim 22,wherein said compound is covalently coupled to said substrate.
 25. Amethod of controlling biofilm formation on a substrate comprising thestep of contacting the compound of claim 16 to said substrate in anamount effective to inhibit biofilm formation or clear a preformedbiofilm.
 26. The method of claim 25, wherein the substrate is selectedfrom the group consisting of a drainpipe, glaze ceramic, porcelain,glass, metal, wood, chrome, plastic, vinyl, and laminate.
 27. A compoundof Formula (V)(a):

wherein: R¹⁸ is selected from the group consisting of: H, hydroxy, acyl,alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl,alkoxy, amino, amide, thiol, sulfone, sulfoxide, oxo, nitro, andcarboxy; R¹⁵, R^(15′), R¹⁶ and R^(16′) are each independently H oralkyl; n=0 to 10, saturated or unsaturated; and p=0 to 10, saturated orunsaturated; or a pharmaceutically acceptable salt or prodrug thereof.28. The compound of claim 27, wherein R¹⁸ is selected from the groupconsisting of: heterocyclo, aryl and heteroaryl.
 29. The compound ofclaim 27, wherein R¹⁸ is a group:

wherein: R⁷ is selected from the group consisting of: H, hydroxy, acyl,alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl,alkoxy, amino, amide, sulfone, sulfoxide, oxo, nitro, carbonyl, andcarboxy; and m=0 to 10, saturated or unsaturated; or a pharmaceuticallyacceptable salt or prodrug thereof.
 30. The compound of claim 27,wherein n=0.
 31. A biofilm preventing, removing or inhibiting coatingcomposition, comprising: (a) a film-forming resin; (b) a solvent thatdisperses said resin; (c) an effective amount of the compound of claim27, wherein said effective amount of said compound prevents or inhibitsthe growth of a biofilm thereon; and (d) optionally, at least onepigment.
 32. A medical device comprising: (a) a medical devicesubstrate; and (b) an effective amount of the compound of claim 27,either coating the substrate, or incorporated into the substrate,wherein said effective amount of said compound prevents or inhibits thegrowth of a biofilm thereon.
 33. The medical device of claim 32, whereinsaid medical device substrate is selected from the group consisting ofstents, fasteners, ports, catheters, scaffolds and grafts.
 34. Themedical device of claim 32, wherein said compound is covalently coupledto said substrate.
 35. A method of controlling biofilm formation on asubstrate comprising the step of contacting the compound of claim 27 tosaid substrate in an amount effective to inhibit biofilm formation orclear a preformed biofilm.
 36. The method of claim 35, wherein thesubstrate is selected from the group consisting of a drainpipe, glazeceramic, porcelain, glass, metal, wood, chrome, plastic, vinyl, andlaminate.